Science.gov

Sample records for 2-dimensional electrophoresis 2-de

  1. Effect of bile salts stress on protein synthesis of Lactobacillus casei Zhang revealed by 2-dimensional gel electrophoresis.

    PubMed

    Wu, R; Sun, Z; Wu, J; Meng, H; Zhang, H

    2010-08-01

    Lactobacillus casei Zhang, isolated from koumiss in Inner Mongolia of China, is known from previous findings to be tolerant to bile salts. Bile salts secreted by mammals act as a natural antibacterial barrier and may serve as a component of innate immunity, as they have limited antagonistic effect against resident microflora. In this work, we compared the growth and protein expression patterns of L. casei Zhang with and without bile salts. Twenty-six proteins were found to be differentially expressed using 2-dimensional gel electrophoresis. Peptide mass fingerprinting was used to identify these proteins. Further verification by using real-time, quantitative reverse transcription-PCR and bioinformatics analysis showed that the implicated pathways are involved with a complex physiological response under bile salts stress, particularly including cell protection (DnaK and GroEL), modifications in cell membranes (NagA, GalU, and PyrD), and key components of central metabolism (PFK, PGM, CysK, LuxS, PepC, and EF-Tu). These results provide insight on the protein expression pattern of L. casei under bile salts stress and offer a new perspective for the molecular mechanisms involved in stress tolerance and adaptation of bacteria. PMID:20655455

  2. Identification and comparative proteomic study of quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry analysis

    PubMed Central

    Hu, Shan; Qiu, Ning; Liu, Yaping; Zhao, Hongyan; Gao, Dan; Song, Rui; Ma, Meihu

    2016-01-01

    A proteomic study of egg white proteins from 2 major poultry species, namely quail (Coturnix coturnix) and duck (Anas platyrhynchos), was performed with comparison to those of chicken (Gallus gallus) through 2-dimensional polyacrylamide gel electrophoresis (2-DE) analysis. By using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS), 29 protein spots representing 10 different kinds of proteins as well as 17 protein spots designating 9 proteins were successfully identified in quail and duck egg white, respectively. This report suggested a closer relationship between quail and chicken egg white proteome patterns, whereas the duck egg white protein distribution on the 2-DE map was more distinct. In duck egg white, some well-known major proteins, such as ovomucoid, clusterin, extracellular fatty acid-binding protein precursor (ex-FABP), and prostaglandin D2 synthase (PG D2 synthase), were not detected, while two major protein spots identified as “deleted in malignant brain tumors 1” protein (DMBT1) and vitellogenin-2 were found specific to duck in the corresponding range on the 2-DE gel map. These interspecies diversities may be associated with the egg white protein functions in cell defense or regulating/supporting the embryonic development to adapt to the inhabiting environment or reproduction demand during long-term evolution. The findings of this work will give insight into the advantages involved in the application on egg white proteins from various egg sources, which may present novel beneficial properties in the food industry or related to human health. PMID:26957635

  3. Identification and comparative proteomic study of quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry analysis.

    PubMed

    Hu, S; Qiu, N; Liu, Y; Zhao, H; Gao, D; Song, R; Ma, M

    2016-05-01

    A proteomic study of egg white proteins from 2 major poultry species, namely quail (Coturnix coturnix) and duck (Anas platyrhynchos), was performed with comparison to those of chicken (Gallus gallus) through 2-dimensional polyacrylamide gel electrophoresis (2-DE) analysis. By using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS), 29 protein spots representing 10 different kinds of proteins as well as 17 protein spots designating 9 proteins were successfully identified in quail and duck egg white, respectively. This report suggested a closer relationship between quail and chicken egg white proteome patterns, whereas the duck egg white protein distribution on the 2-DE map was more distinct. In duck egg white, some well-known major proteins, such as ovomucoid, clusterin, extracellular fatty acid-binding protein precursor (ex-FABP), and prostaglandin D2 synthase (PG D2 synthase), were not detected, while two major protein spots identified as "deleted in malignant brain tumors 1" protein (DMBT1) and vitellogenin-2 were found specific to duck in the corresponding range on the 2-DE gel map. These interspecies diversities may be associated with the egg white protein functions in cell defense or regulating/supporting the embryonic development to adapt to the inhabiting environment or reproduction demand during long-term evolution. The findings of this work will give insight into the advantages involved in the application on egg white proteins from various egg sources, which may present novel beneficial properties in the food industry or related to human health. PMID:26957635

  4. Porcine salivary analysis by 2-dimensional gel electrophoresis in 3 models of acute stress: A pilot study

    PubMed Central

    Fuentes-Rubio, María; Cerón, José J.; de Torre, Carlos; Escribano, Damián; Gutiérrez, Ana M.; Tecles, Fernando

    2014-01-01

    The purpose of this research was to study changes in the salivary proteome of healthy pigs in stressful situations to identify any potential new salivary biomarker of stress. Three groups of animals were subjected to 3 stress models: snaring restraint followed by simulated sampling of vena cava blood; brief transport by road; and restriction of movement in a digestibility cage. Saliva was obtained from each animal before and 15 and 30 min after the induction of stress. The samples from the animals that showed the greatest increase in salivary cortisol concentration were pooled and run on 2-dimensional gels. Coomassie Brilliant Blue R-250 was used for spot detection and mass spectrometry for spot identification. Statistical analyses showed that 2 proteins had significant differences in expression before and after the induction of stress. These proteins were identified as odorant-binding protein and fragments of albumin. Further studies will be necessary to confirm the value of using these proteins as salivary biomarkers of stress in pigs. PMID:24688174

  5. Analysis by enzyme-linked immunosorbent assay and 2-dimensional electrophoresis of haptoglobin in the high-density lipoprotein fraction in cows.

    PubMed

    Kanno, H; Katoh, N

    2001-01-01

    Haptoglobin (Hp) is a hemoglobin (Hb)-binding acute-phase protein. Besides its relevance in inflammation, Hp is involved in the regulation of lipid metabolism. In cattle, in addition to the lipoprotein-deficient fraction, Hp is distributed in high-density lipoprotein (HDL) and very high-density lipoprotein (VHDL) fractions. The purpose of this study was to determine Hp concentrations in the lipoprotein fractions using an enzyme-linked immunosorbent assay (ELISA) based on the affinity with Hb, and also to detect structural differences of HDL Hp from that in the lipoprotein-deficient fraction using 2-dimensional electrophoresis. When purified Hp was used as the antigen for the ELISA, the detection limit was 7.4 ng/ml and linearity was obtained from 14.8 to 475 ng/ml. The correlation coefficient between the ELISA and single radial immunodiffusion was 0.884. The ELISA was shown to be applicable to evaluate Hp concentrations in the lipoprotein fractions. Hp concentrations in the lipoprotein fractions were in the range of 0.94 to 8.77 microg of Hp/ml (n = 4), and concentration ratios were 0.2 to 0.3% of whole serum Hp. Of the lipoprotein fractions, Hp was most abundant in HDL, moderate in VHDL and faint in chylomicrons, the very low-density lipoprotein fraction and low-density lipoprotein fraction. By 2-dimensional electrophoresis, alpha- and beta-chains of serum Hp were each separated into 5 spots, and their isoelectric point (pI) values were from 5.05 to 6.28 in the alpha-chain and from 5.92 to 6.95 in the beta-chain. The pI values of HDL Hp were indistinguishable from those of serum Hp. These results indicate that the ELISA based on the affinity with Hb is useful for evaluating Hp concentrations in lipoprotein fractions, and also suggest that HDL Hp is structurally similar to that in the lipoprotein-deficient fraction. PMID:11217066

  6. Psoriasin, one of several new proteins identified in nasal lavage fluid from allergic and non-allergic individuals using 2-dimensional gel electrophoresis and mass spectrometry

    PubMed Central

    Bryborn, Malin; Adner, Mikael; Cardell, Lars-Olaf

    2005-01-01

    Background Extravasation and luminal entry of plasma occurs continuously in the nose. This process is markedly facilitated in patients with symptomatic allergic rhinitis, resulting in an increased secretion of proteins. Identification of these proteins is an important step in the understanding of the pathological mechanisms in allergic diseases. DNA microarrays have recently made it possible to compare mRNA profiles of lavage fluids from healthy and diseased patients, whereas information on the protein level is still lacking. Methods Nasal lavage fluid was collected from 11 patients with symptomatic allergic rhinitis and 11 healthy volunteers. 2-dimensional gel electrophoresis was used to separate proteins in the lavage fluids. Protein spots were picked from the gels and identified using mass spectrometry and database search. Selected proteins were confirmed with western blot. Results 61 spots were identified, of which 21 were separate proteins. 6 of these proteins (psoriasin, galectin-3, alpha enolase, intersectin-2, Wnt-2B and hypothetical protein MGC33648) had not previously been described in nasal lavage fluids. The levels of psoriasin were markedly down-regulated in allergic individuals. Prolactin-inducible protein was also found to be down-regulated, whereas different fragments of albumin together with Ig gamma 2 chain c region, transthyretin and splice isoform 1 of Wnt-2B were up-regulated among the allergic patients. Conclusion The identification of proteins in nasal lavage fluid with 2-dimensional gelelectrophoresis in combination with mass spectrometry is a novel tool to profile protein expression in allergic rhinitis and it might prove useful in the hunt for new therapeutic targets or diagnostic markers for allergic diseases. Psoriasin is a potent chemotactic factor and its down-regulation during inflammation might be of importance for the outcome of the disease. PMID:16236163

  7. A comparative proteomic study of plasma in feline pancreatitis and pancreatic carcinoma using 2-dimensional gel electrophoresis to identify diagnostic biomarkers: A pilot study

    PubMed Central

    Meachem, Melissa D.; Snead, Elisabeth R.; Kidney, Beverly A.; Jackson, Marion L.; Dickinson, Ryan; Larson, Victoria; Simko, Elemir

    2015-01-01

    While pancreatitis is now recognized as a common ailment in cats, the diagnosis remains challenging due to discordant results and suboptimal sensitivity of ultrasound and specific feline pancreatic lipase (Spec fPL) assay. Pancreatitis also shares similar clinical features with pancreatic carcinoma, a rare but aggressive disease with a grave prognosis. The objective of this pilot study was to compare the plasma proteomes of normal healthy cats (n = 6), cats with pancreatitis (n = 6), and cats with pancreatic carcinoma (n = 6) in order to identify potential new biomarkers of feline pancreatic disease. After plasma protein separation by 2-dimensional gel electrophoresis, protein spots were detected by Coomassie Brilliant Blue G-250 staining and identified by mass spectrometry. Alpha-1-acid glycoprotein (AGP), apolipoprotein-A1 (Apo-A1), and apolipoprotein-A1 precursor (Pre Apo-A1) appeared to be differentially expressed, which suggests the presence of a systemic acute-phase response and alteration of lipid metabolism in cats with pancreatic disease. Future studies involving greater case numbers are needed in order to assess the utility of these proteins as potential biomarkers. More sensitive proteomic techniques may also be helpful in detecting significant but low-abundance proteins. PMID:26130850

  8. Exposures of Sus scrofa to a TASER(®) conducted electrical weapon: no effects on 2-dimensional gel electrophoresis patterns of plasma proteins.

    PubMed

    Jauchem, James R; Cerna, Cesario Z; Lim, Tiffany Y; Seaman, Ronald L

    2014-12-01

    In an earlier study, we found significant changes in red-blood-cell, leukocyte, and platelet counts, and in red-blood-cell membrane proteins, following exposures of anesthetized pigs to a conducted electrical weapon. In the current study, we examined potential changes in plasma proteins [analyzed via two-dimensional gel electrophoresis (2-DGE)] following two 30 s exposures of anesthetized pigs (Sus scrofa) to a TASER (®) C2 conducted electrical weapon. Patterns of proteins, separated by 2-DGE, were consistent and reproducible between animals and between times of sampling. We determined that the blood plasma collection, handling, storage, and processing techniques we used are suitable for swine blood. There were no statistically significant changes in plasma proteins following the conducted-electrical-weapon exposures. Overall gel patterns of fibrinogen were similar to results of other studies of both pigs and humans (in control settings, not exposed to conducted electrical weapons). The lack of significant changes in plasma proteins may be added to the body of evidence regarding relative safety of TASER C2 device exposures. PMID:25319243

  9. Diagonal two-dimensional electrophoresis (D-2DE): a new approach to study the effect of osmotic stress induced by polyethylene glycol in durum wheat (Triticum durum Desf.).

    PubMed

    Kacem, N S; Mauro, S; Muhovski, Y; Delporte, F; Renaut, J; Djekoun, A; Watillon, B

    2016-09-01

    Acclimatization to stress is associated with profound changes in proteome composition. The use of plant cell and tissue culture offers a means to investigate the physiological and biochemical processes involved in the adaptation to osmotic stress. We employed a new proteomic approach to further understand the response of calli to dehydration induced by polyethylene glycol (PEG6000). Calli of three durum wheat genotypes Djenah Khetifa, Oued Zenati and Waha were treated with two concentrations of polyethylene glycol to mimic osmotic stress. Changes in protein relative abundance were analyzed using a new electrophoretic approach named diagonal two-dimensional electrophoresis (D-2DE), combined with mass spectrometry. Total proteins were extracted from 30-day-old calli from three durum wheat genotypes that showed contrasting levels of drought stress tolerance in the field. The combination of one-dimensional electrophoresis and D-2DE gave a specific imprint of the protein extracts under osmotic stress, as well as characterizing and identifying individual target proteins. Of the variously expressed proteins, three were selected (globulin, GAPDH and peroxidase) and further analyzed using qRT-PCR at the transcriptome level in order to compare the results with the proteomic data. Western blot analysis was used to further validate the differences in relative abundance pattern. The proteins identified through this technique provide new insights as to how calli respond to osmotic stress. Our method of study provides an original and relevant approach of analyzing the osmotic-responsive mechanisms at the cellular level of durum wheat with agronomic perspectives. PMID:27317377

  10. Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE).

    PubMed

    Tan, A A; Azman, S N; Abdul Rani, N R; Kua, B C; Sasidharan, S; Kiew, L V; Othman, N; Noordin, R; Chen, Y

    2011-12-01

    There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to perform additional sample preparation steps to improve the quality of the final results, yet without selectively losing the proteins. To address this, we developed a general method that is suitable for diverse sample types based on phenolchloroform extraction method (represented by TRI reagent). This method was found to yield good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae, Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line, bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit, PRO-PREP™ Protein Extraction Solution and lysis buffer. The most useful protocol allows the extraction and separation of a wide diversity of protein samples that is reproducible among repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the highest protein yield as well as the greatest number of total proteins spots count for all type of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of different extraction methods used for each type of sample. PMID:22433892

  11. Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis

    PubMed Central

    Cheng, Hao-Tsai; Sung, Chang-Mu; Pai, Betty Chien-Jung; Liu, Nai-Jen; Chen, Carl PC

    2016-01-01

    Aims. Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established. Methods and Material. The method involved acetone precipitation followed by protein extraction using commercially available 2D Clean-Up kit. The effectiveness was evaluated by 2-dimensional electrophoresis (2DE) profiling quality, including number of protein spots and spot distribution. Results. The total protein of bile fluid in benign biliary disorders was 0.797 ± 0.465 μg/μL. The sample preparation method using acetone precipitation first followed by 2D Clean-Up kit protein extraction resulted in better quality of 2DE gel images in terms of resolution as compared with other sample preparation methods. Using this protocol, we obtained approximately 558 protein spots on the gel images and with better protein spots presentation of haptoglobin, serum albumin, serotransferrin, and transthyretin. Conclusions. Protein samples of bile prepared using acetone precipitation followed by 2D Clean-Up kit exhibited high protein resolution and significant protein profile. This optimized protein preparation protocol can effectively concentrate bile proteins, remove abundant proteins and debris, and yield clear presentation of nonabundant proteins and its isoforms on 2-dimensional electrophoresis gel images. PMID:26966686

  12. Capillary electrophoresis

    SciTech Connect

    Warner, M.

    1988-10-15

    Rapid instrumental methods for performing electrophoretic separations in capillary tubes have recently been developed, making capillary electrophoresis one of the most exciting new techniques available to analytical chemists. This article discusses detection methods, applications, and the future of capillary electrophoresis.

  13. High Resolution Quantitative Proteomics of HeLa Cells Protein Species Using Stable Isotope Labeling with Amino Acids in Cell Culture(SILAC), Two-Dimensional Gel Electrophoresis(2DE) and Nano-Liquid Chromatograpohy Coupled to an LTQ-OrbitrapMass Spectrometer*

    PubMed Central

    Thiede, Bernd; Koehler, Christian J.; Strozynski, Margarita; Treumann, Achim; Stein, Robert; Zimny-Arndt, Ursula; Schmid, Monika; Jungblut, Peter R.

    2013-01-01

    The proteomics field has shifted over recent years from two-dimensional gel electrophoresis (2-DE)-based approaches to SDS-PAGE or gel-free workflows because of the tremendous developments in isotopic labeling techniques, nano-liquid chromatography, and high-resolution mass spectrometry. However, 2-DE still offers the highest resolution in protein separation. Therefore, we combined stable isotope labeling with amino acids in cell culture of controls and apoptotic HeLa cells with 2-DE and the subsequent analysis of tryptic peptides via nano-liquid chromatography coupled to an LTQ-Orbitrap mass spectrometer to obtain quantitative data using the methods with the highest resolving power on all levels of the proteomics workflow. More than 1,200 proteins with more than 2,700 protein species were identified and quantified from 816 Coomassie Brilliant Blue G-250 stained 2-DE spots. About half of the proteins were identified and quantified only in single 2-DE spots. The majority of spots revealed one to five proteins; however, in one 2-DE spot, up to 23 proteins were identified. Only half of the 2-DE spots represented a dominant protein with more than 90% of the whole protein amount. Consequently, quantification based on staining intensities in 2-DE gels would in approximately half of the spots be imprecise, and minor components could not be quantified. These problems are circumvented by quantification using stable isotope labeling with amino acids in cell culture. Despite challenges, as shown in detail for lamin A/C and vimentin, the quantitative changes of protein species can be detected. The combination of 2-DE with high-resolution nano-liquid chromatography-mass spectrometry allowed us to identify proteomic changes in apoptotic cells that would be unobservable using any of the other previously employed proteomic workflows. PMID:23033477

  14. Bargain Electrophoresis.

    ERIC Educational Resources Information Center

    Maderia, Vitor M. C.; Pires, Euclides M. V.

    1986-01-01

    Discusses the value of electrophoresis in the fields of protein chemistry and biochemistry. Describes how to build an inexpensive electrophoresis setup for use in either research or teaching activities. Details the construction of both the separating device and the power supply. (TW)

  15. Tuber borchii fruit body: 2-dimensional profile and protein identification.

    PubMed

    Pierleoni, Raffaella; Buffalini, Michele; Vallorani, Luciana; Guidi, Chiara; Zeppa, Sabrina; Sacconi, Cinzia; Pucci, Piero; Amoresano, Angela; Casbarra, Annarita; Stocchi, Vilberto

    2004-04-01

    The formation of the fruit body represents the final phase of the ectomycorrhizal fungus T. borchii life cycle. Very little is known concerning the molecular and biochemical processes involved in the fructification phase. 2-DE maps of unripe and ripe ascocarps revealed different protein expression levels and the comparison of the electropherograms led to the identification of specific proteins for each developmental phase. Associating micropreparative 2-DE to microchemical approaches, such as N-terminal sequencing and 2-D gel-electrophoresis mass-spectrometry, proteins playing pivotal roles in truffle physiology were identified. PMID:15081280

  16. Electrophoresis technology

    NASA Technical Reports Server (NTRS)

    Snyder, R. S.

    1985-01-01

    A new high resolution apparatus designed for space was built as a laboratory prototype. Using a moving wall with a low zeta potential coating, the major sources of flow distortion for an electrophoretic sample stream are removed. Highly resolved fractions, however, will only be produced in space because of the sensitivity of this chamber to buoyancy-induced convection in the laboratory. The second and third flights of the McDonnell Douglas Astronautics Corporation continuous flow electrophoresis system carried samples developed at MSFC intended to evaluate the broad capabilities of free flow electrophoresis in a reduced gravity environment. Biological model materials, hemoglobin and polystyrene latex microspheres, were selected because of their past use as electrophoresis standards and as visible markers for fluid flow due to electroosmosis, spacecraft acceleration or other factors. The dependence of the separation resolution on the properties of the sample and its suspension solution was assessed.

  17. Simulating Electrophoresis.

    ERIC Educational Resources Information Center

    Moertel, Cheryl; Frutiger, Bruce

    1996-01-01

    Describes a DNA fingerprinting simulation that uses vegetable food coloring and plastic food containers instead of DNA and expensive gel electrophoresis chambers. Allows students to decipher unknown combinations of dyes in a method similar to that used to decipher samples of DNA in DNA fingerprint techniques. (JRH)

  18. Electrophoresis. [in microgravity environment

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1977-01-01

    Ground-based techniques for electrophoresis take account of the need either to circumvent the effects of gravity to prevent convection, or to use gravity for fluid stabilization through artificial density gradients. The microgravity environments of orbiting spacecraft provides a new alternative for electrophoresis by avoiding the need for either of these two approaches. The paper presents some theoretical considerations concerning electrophoresis, examines certain experimental techniques (zone and high density gel electrophoresis, isoelectric focusing and isotachophoresis), and examines the electrophoresis of living cells.

  19. Kidney cell electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P.

    1980-01-01

    The following aspects of kidney cell electrophoresis are discussed: (1) the development and testing of electrophoresis solutions; (2) optimization of freezing and thawing; (3) procedures for evaluation of separated kidney cells; and (4) electrophoretic mobility characterization of kidney cells.

  20. Kidney cell electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P.

    1979-01-01

    A kidney cell electrophoresis technique is described in four parts: (1) the development and testing of electrophoresis solutions; (2) optimization of freezing and thawing; (3) procedures for evaluation of separated kidney cells; and (4) electrophoretic mobility characteristics of kidney cells.

  1. Kidney cell electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P. W.

    1985-01-01

    Tasks were undertaken in support of two objectives. They are: (1) to carry out electrophoresis experiments on cells in microgravity; and (2) assess the feasibility of using purified kidney cells from embryonic kidney cultures as a source of important cell products. Investigations were carried out in the following areas: (1) ground based electrophoresis technology; (2) cell culture technology; (3) electrophoresis of cells; (4) urokinase assay research; (5) zero-g electrophoresis; and (6) flow cytometry.

  2. Comparative proteomics and difference gel electrophoresis.

    PubMed

    Minden, Jonathan

    2007-12-01

    The goal of comparative proteomics is to analyze proteome changes in response to development, disease, or environment. This is a two-step process in which proteins within cellular extracts are first fractionated to reduce sample complexity, and then the proteins are identified by mass spectrometry. Two-dimensional electrophoresis (2DE) is the long-time standard for protein separation, but it has suffered from poor reproducibility and limited sensitivity. Difference gel electrophoresis (DIGE), in which two protein samples are separately labeled with different fluorescent dyes and then co-electrophoresed on the same 2DE gel, was developed to overcome the reproducibility and sensitivity limitations. In this essay, I discuss the principles of comparative proteomics and the development of DIGE. PMID:18251249

  3. Kidney Cell Electrophoresis

    NASA Technical Reports Server (NTRS)

    Todd, P.

    1985-01-01

    Materials and procedures for microgravity electrophoresis of living human embryonic kidney cells were evaluated, ground support in the form of analytical cell electrophoresis and flow cytometry was provided and cells returned from space flight were analyzed. Preflight culture media, electrophoresis buffer, fraction collection media, temperature profiles, and urokinase assay procedures were tested prior to flight. Electrophoretic mobility distributions of aliquots of the cell population to be fractionated in flight were obtained. The protocol established and utilized is given.

  4. Electrophoresis for biological production

    NASA Technical Reports Server (NTRS)

    Mccreight, L. R.

    1977-01-01

    Preparative electrophoresis may provide a unique method for meeting ever more stringent purity requirements. Prolonged near zero gravity in space may permit the operation of preparative electrophoresis equipment with 100 times greater throughput than is currently available. Some experiments with influenza Virus Antigen, Erythropoietin and Antihemophaliac Factor, along with process and economic projections, are briefly reviewed.

  5. Electrophoresis of biological materials

    NASA Technical Reports Server (NTRS)

    1975-01-01

    The selection of biological products was studied for electrophoresis in space. Free flow electrophoresis, isoelectric focusing, and isotachophoresis are described. The candidates discussed include: immunoglobulins and gamma globulins; isolated islet of langerhans from pancreas; bone marrow; tumor cells; kidney cells, cryoprecipitate; and column separated cultures.

  6. Automatic multiple applicator electrophoresis

    NASA Technical Reports Server (NTRS)

    Grunbaum, B. W.

    1977-01-01

    Easy-to-use, economical device permits electrophoresis on all known supporting media. System includes automatic multiple-sample applicator, sample holder, and electrophoresis apparatus. System has potential applicability to fields of taxonomy, immunology, and genetics. Apparatus is also used for electrofocusing.

  7. Simulation of Two Dimensional Electrophoresis and Tandem Mass Spectrometry for Teaching Proteomics

    ERIC Educational Resources Information Center

    Fisher, Amanda; Sekera, Emily; Payne, Jill; Craig, Paul

    2012-01-01

    In proteomics, complex mixtures of proteins are separated (usually by chromatography or electrophoresis) and identified by mass spectrometry. We have created 2DE Tandem MS, a computer program designed for use in the biochemistry, proteomics, or bioinformatics classroom. It contains two simulations--2D electrophoresis and tandem mass spectrometry.…

  8. Basics and recent advances of two dimensional- polyacrylamide gel electrophoresis

    PubMed Central

    2014-01-01

    Gel- based proteomics is one of the most versatile methods for fractionating protein complexes. Among these methods, two dimensional- polyacrylamide gel electrophoresis (2-DE) represents a mainstay orthogonal approach, which is popularly used to simultaneously fractionate, identify, and quantify proteins when coupled with mass spectrometric identification or other immunological tests. Although 2-DE was first introduced more than three decades ago, several challenges and limitations to its utility still exist. This review discusses the principles of 2-DE as well as both recent methodological advances and new applications. PMID:24735559

  9. (1+2)-dimensional strongly nonlocal solitons

    SciTech Connect

    Ouyang Shigen; Guo Qi

    2007-11-15

    Approximate solutions of (1+2)-dimensional strongly nonlocal solitons (SNSs) are presented. It is shown that the power of a SNS in a nematic liquid crystal is in direct proportion to the second power of the degree of nonlocality, the power of a SNS in a nonlocal medium with a logarithmic nonlocal response is in inverse proportion to the second power of its beamwidth, and the power of a SNS in a nonlocal medium with an sth-power decay nonlocal response is in direct proportion to the (s+2)th power of the degree of nonlocality.

  10. Preparative electrophoresis experiment design

    NASA Technical Reports Server (NTRS)

    Thiehler, A.

    1972-01-01

    A multifaceted study supporting the NASA programs to develop a space electrophoresis capability has been conducted. The study involved principally the technique of continuous free electrophoresis. It comprised a critical review of the art, study of new techniques for enhancing resolution and stability, and construction and initial testing of a high resolution cell. The effort resulted in a significant advance in free electrophoresis technique. It has provided also a much improved base for developments exploiting the added advantages of a zero-gravity environment.

  11. Anisotropic 2-dimensional Robin Hood model

    NASA Astrophysics Data System (ADS)

    Buldyrev, Sergey; Cwilich, Gabriel; Zypman, Fredy

    2009-03-01

    We have considered the Robin Hood model introduced by Zaitsev[1] to discuss flux creep and depinning of interfaces in a two dimensional system. Although the model has been studied extensively analytically in 1-d [2], its scaling laws have been verified numerically only in that case. Recent work suggest that its properties might be important to understand surface friction[3], where its 2-dimensional properties are important. We show that in the 2-dimensional case scaling laws can be found provided one considers carefully the anisotropy of the model, and different ways of introducing that anisotropy lead to different exponents and scaling laws, in analogy with directed percolation, with which this model is closely related[4]. We show that breaking the rotational symmetry between the x and y axes does not change the scaling properties of the model, but the introduction of a preferential direction of accretion (``robbing'' in the language of the model) leads to new scaling exponents. [1] S.I.Zaitsev, Physica A189, 411 (1992) [2] M. Pacuzki, S. Maslov and P.Bak, Phys Rev. E53, 414 (1996) [3] S. Buldyrev, J. Ferrante and F. Zypman Phys. Rev E64, 066110 (2006) [4] G. Odor, Rev. Mod. Phys. 76, 663 (2004) .

  12. Serum globulin electrophoresis

    MedlinePlus

    ... may indicate: Acute infection Bone marrow cancer called multiple myeloma Chronic inflammatory disease (for example, rheumatoid arthritis and ... test Hemoglobin Hyperimmunization Immunoelectrophoresis - ... electrophoresis - serum Rheumatoid arthritis Systemic lupus erythematosus ...

  13. Recent advances in preparative electrophoresis

    NASA Technical Reports Server (NTRS)

    Mosher, Richard A.; Thormann, Wolfgang; Egen, Ned B.; Couasnon, Pascal; Sammons, David W.

    1987-01-01

    Various approaches for preparative electrophoresis, and three new instruments for preparative electrophoresis are discussed. Consideration is given to isoelectric focusing, isotachophoresis, and zone electrophoresis, three gel-based electrophoresis methods. The design, functions, and performance of the Elphor VaP 21 device of Hannig (1982), the shear-stabilized BIOSTREAM separator of Thompson (1983), and the recycling isoelectric focusing device are described.

  14. DNA ELECTROPHORESIS AT SURFACES

    SciTech Connect

    RAFAILOVICH, MIRIAM; SOKOLOV, JONATHAN; GERSAPPE, DILIP

    2003-09-01

    During this year we performed two major projects: I. We developed a detailed theoretical model which complements our experiments on surface DNA electrophoresis. We found that it was possible to enhance the separation of DNA chains by imposing a chemical nanoscale pattern on the surface. This approach utilized the surface interaction effect of the DNA chains with the substrate and is a refinement to our previous method in which DNA chains were separated on homogeneous flat surfaces. By introducing the nano-patterns on the surface, the conformational changes of DNA chains of different lengths can be amplified, which results in the different friction strengths with the substrate surface. Our results also show that, when compared to the DNA electrophoresis performed on homogeneous flat surfaces, nanopatterned surfaces offer a larger window in choosing different surface interactions to achieve separation. II. In collaboration with a large international manufacturer of skin care products we also embarked on a project involving photo toxicity of titanium dioxide nanoparticles, which are a key ingredient in sunscreen and cosmetic lotions. The results clearly implicated the nanoparticles in catalyzing damage to chromosomal DNA. We then used this knowledge to develop a polymer/anti-oxidant coating which prevented the photocatalytic reaction on DNA while still retaining the UV absorptive properties of the nanoparticles. The standard gel electrophoresis was not sufficient in determining the extent of the DNA damage. The conclusions of this study were based predominantly on analysis obtained with the surface electrophoresis method.

  15. Fraction collector for electrophoresis

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1977-01-01

    Rotating-tube electrophoresis apparatus employs rotating jet of eluting buffer to reduce effects of convection during separation. Designed for separation of microorganisms and biological species, system combines gravity/gradient compensating of lumen with buffer flush at fraction outlet to increase separation efficiency.

  16. Analysis of electrophoresis performance

    NASA Technical Reports Server (NTRS)

    Roberts, G. O.

    1984-01-01

    The SAMPLE computer code models electrophoresis separation in a wide range of conditions. Results are included for steady three dimensional continuous flow electrophoresis (CFE), time dependent gel and acetate film experiments in one or two dimensions and isoelectric focusing in one dimension. The code evolves N two dimensional radical concentration distributions in time, or distance down a CFE chamber. For each time or distance increment, there are six stages, successively obtaining the pH distribution, the corresponding degrees of ionization for each radical, the conductivity, the electric field and current distribution, and the flux components in each direction for each separate radical. The final stage is to update the radical concentrations. The model formulation for ion motion in an electric field ignores activity effects, and is valid only for low concentrations; for larger concentrations the conductivity is, therefore, also invalid.

  17. Segmented field OFFGEL® electrophoresis.

    PubMed

    Tobolkina, Elena; Cortés-Salazar, Fernando; Momotenko, Dmitry; Maillard, Julien; Girault, Hubert H

    2012-11-01

    A multielectrode setup for protein OFFGEL electrophoresis that significantly improves protein separation efficiency has been developed. Here, the electric field is applied by segments between seven electrodes connected in series to six independent power supplies. The aim of this strategy is to distribute evenly the electric field along the multiwell system, and as a consequence to enhance electrophoresis in terms of separation time, resolution, and protein collection efficiency, while minimizing the overall potential difference and therefore the Joule heating. The performances were compared to a standard two-electrode setup for OFFGEL fractionation of a protein mixture, using UV-Vis spectroscopy for quantification and MALDI-MS for identification. The electrophoretic separation process was simulated, and optimized by solving the time-dependent Nernst-Planck differential equation. PMID:23086720

  18. Happy bicentennial, electrophoresis!

    PubMed

    Righetti, Pier Giorgio

    2009-12-01

    A short survey of electrophoresis and a celebration of its bicentennial, with some remarkable mementos and a list of books that shaped the field. Where one also learns of a secret production plant with a huge-scale electrophoretic apparatus for skimming of latex from Hevea brasiliensis and keeping the wheels of the Ally Army running during World War II. And of cyber (mammoth) 2D gels of 1.5 x 1 m in size accommodating >12,000 spots. PMID:19938305

  19. Preparative electrophoresis for space

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H.; Snyder, Robert S.

    1988-01-01

    A premise of continuous flow electrophoresis is that removal of buoyance-induced thermal convection caused by axial and lateral temperature gradients results in ideal performance of these instruments in space. Although these gravity dependent phenomena disturb the rectilinear flow in the separation chamber when high voltage gradients or thick chamber are used, distortion of the injected sample stream due to electrodynamic effects cause major broadening of the separated bands. The electrophoresis separation process is simple, however flow local to the sample filament produced by the applied electric field were not considered. These electrohydrodynamic flows distort the sample stream and limit the separation. Also, electroosmosis and viscous flow combine to further distort the process. A moving wall concept is being proposed for space which will eliminate and control the disturbances. The moving wall entrains the fluid to move as a rigid body and produces a constant residence time for all samples distributed across the chamber thickness. The moving wall electrophoresis chamber can only be operated in space because there is no viscous flow in the chamber to stabilize against thermal convection.

  20. Preparative electrophoresis for space

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H.; Snyder, Robert S.

    1987-01-01

    A premise of continuous flow electrophoresis is that removal of buoyancy-induced thermal convection caused by axial and lateral temperature gradients results in ideal performance of these instruments in space. Although these gravity dependent phenomena disturb the rectilinear flow in the separation chamber when high voltage gradients or thick chambers are used, distortion of the injected sample stream due to electrohydrodynamic effects cause major broadening of the separated bands. The electrophoresis separation process is simple, however flow local to the sample filament produced by the applied electric field have not been considered. These electrohydrodynamic flows distort the sample stream and limit the separation. Also, electroosmosis and viscous flow combine to further distort the process. A moving wall concept is being proposed for space which will eliminate and control the disturbances. The moving wall entrains the fluid to move as a rigid body and produces a constant residence time for all samples distributed across the chamber thickness. The moving wall electrophoresis chamber can only be operated in space because there is no viscous flow in the chamber to stabilize against thermal convection.

  1. Generalized elastica on 2-dimensional de Sitter space S12

    NASA Astrophysics Data System (ADS)

    Huang, Rongpei; Yu, Junyan

    2016-02-01

    In this paper, the extremals of curvature energy actions on non-null regular curves in 2-dimensional de Sitter space are studied. We completely solve the Euler-Lagrange equation by quadratures. By using the Killing field, we construct three special coordinate systems and express the generalized elastica in 2-dimensional de Sitter space S12 by integral explicitly.

  2. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, Edward S.; Li, Qingbo; Lu, Xiandan

    1998-04-21

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

  3. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, Edward S.; Chang, Huan-Tsang; Fung, Eliza N.; Li, Qingbo; Lu, Xiandan

    1996-12-10

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

  4. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, E.S.; Li, Q.; Lu, X.

    1998-04-21

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

  5. Multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, E.S.; Chang, H.T.; Fung, E.N.; Li, Q.; Lu, X.

    1996-12-10

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

  6. Spot Matching of 2-DE Images Using Distance, Intensity, and Pattern Information.

    PubMed

    Xin, Hua-Mei; Zhu, Yuemin

    2016-01-01

    The analysis of a large number of two-dimensional gel electrophoresis (2-DE) images requires developing automatic methods. In such analyses, spot matching plays a fundamental role, in particular for the identification of proteins. We describe a simple and accurate method which allows to automatically and accurately match spots in 2-DE images. The method consists of simultaneously exploiting the distance between the spots, their intensity, and the pattern formed by their spatial configuration. PMID:26611412

  7. Two-dimensional electrophoresis of Arenicola marina extracellular hemoglobin: separation of chains with identical molecular mass but different isoelectric point.

    PubMed

    Slitine, F E; Toulmond, A

    1991-01-01

    1. On the basis of their molecular masses, four types of polypeptides (A, B, C, D) were obtained by SDS-PAGE of the extracellular hemoglobin of the polychaete annelid Arenicola marina. 2. On 2-dimensional polyacrylamide gel electrophoresis, the erythrocruorin dissociated into six different types of polypeptide chains; A1, A2, B1, B2, C and D. 3. A1 and B1 migrate in 2-dimensional electrophoresis at the same position as alpha and beta chains of human hemoglobin. PMID:1814687

  8. Apparatus for electrophoresis separation

    DOEpatents

    Anderson, Norman L.

    1978-01-01

    An apparatus is disclosed for simultaneously performing electrophoresis separations on a plurality of slab gels containing samples of protein, protein subunits or nucleic acids. A reservoir of buffer solution is divided into three compartments by two parallel partitions having vertical slots spaced along their length. A sheet of flexible, electrically insulative material is attached to each partition and is provided with vertical slits aligned with the slots. Slab-gel holders are received within the slots with the flexible material folded outwardly as flaps from the slits to overlay portions of the holder surfaces and thereby act as electrical and liquid seals. An elongated, spaghetti-like gel containing a sample of specimen that was previously separated by isoelectric focusing techniques is vertically positioned along a marginal edge portion of the slab gel. On application of an electrical potential between the two outer chambers of buffer solution, a second dimensional electrophoresis separation in accordance with molecular weight occurs as the specimen molecules migrate across the slab gel.

  9. Fluorescence Detection In Electrophoresis

    NASA Astrophysics Data System (ADS)

    Swarner, Susan

    1988-04-01

    Fluorescence detection is in common usage in forensic science laboratories for the visualization of three enzyme markers. The fluorogenic substrates, 4-methylumbelliferyl phosphate, 4-methylutbel-liveryl acetate, and fluorecein diacetate, are acted upon by the enzymes Erythrocyte Acid Phospha, tase, Esterase-D, and Carbonic Anhydrase-III, respectively, to produce compounds visible to the analyst when viewed with transmitted UV light at 365 nm. Additionally, the choice of fluorogenic corn, pounds may help detect a specific enzyme from a related enzyme. One of the responsibilities of a forensic science laboratory may be the analysis of blood for genetically controlled polymorphic enzymes and protein markers. The genetic markers are said to be polymorphic because each exhibits types which can be differentiated and allows for the inclusion or exclusion of possible-donors of the blood. Each genetic marker can be separated into these recognizable types by electrophoresis, a technique which separates compounds based on electrical charges. Electrophoresis is conducted by placing a portion or extract of each bloodstain into a support medium which will conduct electricity. This is known as a plate or membrane. By controlling the pH of the buffer and the potential that is applied to the plate, the analyst can achieve separation of the types within an enzyme marker. The types appear as differing patterns of bands. Once the bloodstain has been subjected to electrophoresis, the enzymes must be visualized. This is generally best accomplished by using the specific activity of the enzyme. For the enzymes described in the present work, the visualization is performed by over-layering the plate with a piece of filter paper that 'has been saturated with the appropriate non-fluorescent substrate and buffer. The bands of enzyme, which is now in discrete patterns, will act upon the non-fluorescent substrate to create a fluorescent compound. The plate is then viewed with transmitted UV

  10. Static continuous electrophoresis device

    NASA Technical Reports Server (NTRS)

    Rhodes, P. H. (Inventor)

    1982-01-01

    An apparatus is disclosed for carrying out a moving wall type electrophoresis process for separation of cellular particles. The apparatus includes a water-tight housing containing an electrolytic buffer solution. A separation chamber in the housing is defined by spaced opposed moving walls and spaced opposed side walls. Substrate assemblies, which support the moving wall include vacuum ports for positively sealing the moving walls against the substrate walls. Several suction conduits communicate with the suction ports and are arranged in the form of valleys in a grid plate. The raised land portion of the grid plat supports the substrate walls against deformation inwardly under suction. A cooling chamber is carried on the back side of plate. The apparatus also has tensioner means including roller and adjustment screws for maintaining the belts in position and a drive arrangement including an electric motor with a gear affixed to its output shaft. Electrode assemblies are disposed to provide the required electric field.

  11. Electrophoresis experiment for space

    NASA Technical Reports Server (NTRS)

    Vanderhoff, J. W.; Micale, F. J.

    1976-01-01

    The Apollo 16 electrophoresis experiment was analyzed, demonstrating that the separation of the two different-size monodisperse latexes did indeed take place, but that the separation was obscured by the pronounced electroosmotic flow of the liquid medium. The results of this experiment, however, were dramatic since it is impossible to carry out a similar separation on earth. It can be stated unequivocally from this experiment that any electrophoretic separation will be enhanced under microgravity conditions. The only question is the degree of this enhancement, which can be expected to vary from one experimental technique to another. The low-electroosmotic-mobility coating (Z6040-MC) developed under this program was found to be suitable for a free-fluid electrophoretic separation such as the experiment designed for the ASTP flight. The problem with this coating, however, is that its permanency is limited because of the slow desorption of the methylcellulose from the coated surface.

  12. Kidney cell electrophoresis, continuing task

    NASA Technical Reports Server (NTRS)

    Todd, P. W.

    1985-01-01

    Materials and procedures for microgravity electrophoresis of living human embryonic kidney cells were evaluated to provide ground support in the form of analytical cell electrophoresis and flow cytometry. Preflight culture media, electrophoresis buffer, fraction collection media, temperature profiles, and urokinase assay procedures were tested prior to flight. Electrophoretic mobility distributions of aliquots of the cell population to be fractionated in flight were obtained. Cells were prepared in suspension prior to flight in electrophoresis buffer and 10% calf serum. Electrophoretic separation proceeded in electrophoresis buffer without serum in the Continuous Flow Electrophoretic Separator, and fractions were collected into sample bags containing culture medium and concentrated serum. Fractions that yielded enough progeny cells were analyzed for morphology and electrophoretic mobility distributions. It is noted that the lowest mobility fraction studied produced higher mobility progeny while the other fractions produced progeny cells with mobilities related to the fractions from which they were collected.

  13. Electrophoresis demonstration on Apollo 16

    NASA Technical Reports Server (NTRS)

    Snyder, R. S.

    1972-01-01

    Free fluid electrophoresis, a process used to separate particulate species according to surface charge, size, or shape was suggested as a promising technique to utilize the near zero gravity condition of space. Fluid electrophoresis on earth is disturbed by gravity-induced thermal convection and sedimentation. An apparatus was developed to demonstrate the principle and possible problems of electrophoresis on Apollo 14 and the separation boundary between red and blue dye was photographed in space. The basic operating elements of the Apollo 14 unit were used for a second flight demonstration on Apollo 16. Polystyrene latex particles of two different sizes were used to simulate the electrophoresis of large biological particles. The particle bands in space were extremely stable compared to ground operation because convection in the fluid was negligible. Electrophoresis of the polystyrene latex particle groups according to size was accomplished although electro-osmosis in the flight apparatus prevented the clear separation of two particle bands.

  14. Procedures for two-dimensional electrophoresis of proteins

    SciTech Connect

    Tollaksen, S.L.; Giometti, C.S.

    1996-10-01

    High-resolution two-dimensional gel electrophoresis (2DE) of proteins, using isoelectric focusing in the first dimension and sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) in the second, was first described in 1975. In the 20 years since those publications, numerous modifications of the original method have evolved. The ISO-DALT system of 2DE is a high-throughput approach that has stood the test of time. The problem of casting many isoelectric focusing gels and SDS-PAGE slab gels (up to 20) in a reproducible manner has been solved by the use of the techniques and equipment described in this manual. The ISO-DALT system of two-dimensional gel electrophoresis originated in the late 1970s and has been modified many times to improve its high-resolution, high-throughput capabilities. This report provides the detailed procedures used with the current ISO-DALT system to prepare, run, stain, and photograph two-dimensional gels for protein analysis.

  15. Binary Oscillatory Crossflow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Molloy, Richard F.; Gallagher, Christopher T.; Leighton, David T., Jr.

    1997-01-01

    Electrophoresis has long been recognized as an effective analytic technique for the separation of proteins and other charged species, however attempts at scaling up to accommodate commercial volumes have met with limited success. In this report we describe a novel electrophoretic separation technique - Binary Oscillatory Crossflow Electrophoresis (BOCE). Numerical simulations indicate that the technique has the potential for preparative scale throughputs with high resolution, while simultaneously avoiding many problems common to conventional electrophoresis. The technique utilizes the interaction of an oscillatory electric field and a transverse oscillatory shear flow to create an active binary filter for the separation of charged protein species. An oscillatory electric field is applied across the narrow gap of a rectangular channel inducing a periodic motion of charged protein species. The amplitude of this motion depends on the dimensionless electrophoretic mobility, alpha = E(sub o)mu/(omega)d, where E(sub o) is the amplitude of the electric field oscillations, mu is the dimensional mobility, omega is the angular frequency of oscillation and d is the channel gap width. An oscillatory shear flow is induced along the length of the channel resulting in the separation of species with different mobilities. We present a model that predicts the oscillatory behavior of charged species and allows estimation of both the magnitude of the induced convective velocity and the effective diffusivity as a function of a in infinitely long channels. Numerical results indicate that in addition to the mobility dependence, the steady state behavior of solute species may be strongly affected by oscillating fluid into and out of the active electric field region at the ends of the cell. The effect is most pronounced using time dependent shear flows of the same frequency (cos((omega)t)) flow mode) as the electric field oscillations. Under such conditions, experiments indicate that

  16. Polyacrylamide gel electrophoresis.

    PubMed

    Chrambach, A; Rodbard, D

    1971-04-30

    Polyacrylamide gel electrophoresis (PAGE) provides a versatile, gentle, high resolution method for fractionation and physical-chemical characterization of molecules on the basis of size, conformation, and net charge. The polymerization reaction can be rigorously controlled to provide uniform gels of reproducible, measurable pore size over a wide range. This makes it possible to obtain reproducible relative mobility (Rf) values as physical-chemical constants. Application and extension of Ogston's (random fiber) model for a gel allows for calculation of molecular volume, surface area, or radius, free mobility, and valence from RJ measurements at several gel concentrations, to calculate gel concentration for optimal resolution, and to predict behavior of macromolecules on gel gradients by computerized methods. Extension of classical moving boundary theory has been used to generate multiphasic buffer systems (providing selective stacking, unstacking, restacking, and preparative steady-state-stacking) with known operating characteristics for any pH at 0 degrees and 25 degrees C. A general strategy for isolation of macromolecules and for macromolecular mapping has been developed. Preparative scale PAGE is operational for milligram loads and feasible for gram quantities. PMID:4927678

  17. Microchamber integration unifies distinct separation modes for two-dimensional electrophoresis

    PubMed Central

    Tentori, Augusto M.; Hughes, Alex J.; Herr, Amy E.

    2013-01-01

    By combining isoelectric focusing (IEF) with subsequent gel electrophoresis, two-dimensional electrophoresis (2DE) affords more specific characterization of proteins than each constituent unit separation. In a new approach to integrating the two assay dimensions in a microscope slide-sized glass device, we introduce microfluidic 2DE using photopatterned polyacrylamide (PA) gel elements housed in a millimeter-scale, 20 micron-deep chamber. The microchamber minimizes information loss inherent to channel network architectures commonly used for microfluidic 2DE. To define the IEF axis along a ‘lane’ at the top of the chamber, we used free solution carrier ampholytes and immobilized acrylamido buffers in the PA gels. This approach yielded high resolution (0.1 pH units) and rapid (<20 min) IEF. Next, protein transfer to the second dimension was accomplished using chemical mobilization perpendicular to the IEF axis. Mobilization drove focused proteins off the IEF lane and into a region for protein gel electrophoresis. Using fluorescently labeled proteins, we observed transfer induced band broadening factors ~7.5-fold lower than those observed in microchannel networks. Both native polyacrylamide gel electrophoresis (PAGE) and pore-limit electrophoresis (PLE) were studied as the second assay dimension and completed in <15 min. PLE yields protein molecular mass information without the need for ionic surfactant or reducing agents, simplifying device design and operation. Microchamber-based 2DE unifies two independent separation dimensions in a single device with minimal transfer-associated information losses. Peak capacities for the total assay ranged from 256 to 35 with <1 hr assay duration. The rapid microchamber 2DE assay has the potential to bridge an existing gap in targeted proteomics for protein biomarker validation and systems biology that may complement recent innovation in mass spectrometry. PMID:23565932

  18. Microchamber integration unifies distinct separation modes for two-dimensional electrophoresis.

    PubMed

    Tentori, Augusto M; Hughes, Alex J; Herr, Amy E

    2013-05-01

    By combining isoelectric focusing (IEF) with subsequent gel electrophoresis, two-dimensional electrophoresis (2DE) affords more specific characterization of proteins than each constituent unit separation. In a new approach to integrating the two assay dimensions in a microscope slide-sized glass device, we introduce microfluidic 2DE using photopatterned polyacrylamide (PA) gel elements housed in a millimeter-scale, 20-μm-deep chamber. The microchamber minimizes information loss inherent to channel network architectures commonly used for microfluidic 2DE. To define the IEF axis along a "lane" at the top of the chamber, we used free solution carrier ampholytes and immobilized acrylamido buffers in the PA gels. This approach yielded high-resolution (0.1 pH unit) and rapid (<20 min) IEF. Next, protein transfer to the second dimension was accomplished by chemical mobilization perpendicular to the IEF axis. Mobilization drove focused proteins off the IEF lane and into a region for protein gel electrophoresis. Using fluorescently labeled proteins, we observed transfer-induced band broadening factors ~7.5-fold lower than those observed in microchannel networks. Both native polyacrylamide gel electrophoresis (PAGE) and pore-limit electrophoresis (PLE) were studied as the second assay dimension and completed in <15 min. PLE yields protein molecular mass information without the need for ionic surfactant or reducing agents, simplifying device design and operation. Microchamber-based 2DE unifies two independent separation dimensions in a single device with minimal transfer-associated information losses. Peak capacities for the total assay ranged from 256 to 35 with <1 h assay duration. The rapid microchamber 2DE assay has the potential to bridge an existing gap in targeted proteomics for protein biomarker validation and systems biology that may complement recent innovation in mass spectrometry. PMID:23565932

  19. Copolymers For Capillary Gel Electrophoresis

    DOEpatents

    Liu, Changsheng; Li, Qingbo

    2005-08-09

    This invention relates to an electrophoresis separation medium having a gel matrix of at least one random, linear copolymer comprising a primary comonomer and at least one secondary comonomer, wherein the comonomers are randomly distributed along the copolymer chain. The primary comonomer is an acrylamide or an acrylamide derivative that provides the primary physical, chemical, and sieving properties of the gel matrix. The at least one secondary comonomer imparts an inherent physical, chemical, or sieving property to the copolymer chain. The primary and secondary comonomers are present in a ratio sufficient to induce desired properties that optimize electrophoresis performance. The invention also relates to a method of separating a mixture of biological molecules using this gel matrix, a method of preparing the novel electrophoresis separation medium, and a capillary tube filled with the electrophoresis separation medium.

  20. DNA typing by capillary electrophoresis

    SciTech Connect

    Zhang, N.

    1997-10-08

    Capillary electrophoresis is becoming more and more important in nucleic acid analysis including DNA sequencing, typing and disease gene measurements. This work summarized the background of DNA typing. The recent development of capillary electrophoresis was also discussed. The second part of the thesis showed the principle of DNA typing based on using the allelic ladder as the absolute standard ladder in capillary electrophoresis system. Future work will be focused on demonstrating DNA typing on multiplex loci and examples of disease diagnosis in the on-line format of PCR-CE. Also capillary array electrophoresis system should allow high throughput, fast speed DNA typing. Only the introduction and conclusions for this report are available here. A reprint was removed for separate processing.

  1. Biomedical applications of capillary electrophoresis

    NASA Astrophysics Data System (ADS)

    Kartsova, L. A.; Bessonova, E. A.

    2015-08-01

    The review deals with modern analytical approaches used in capillary electrophoresis for solving medical and biological problems: search for biomarkers of various diseases and rapid diagnosis based on characteristic profiles of biologically active compounds by capillary electrophoresis with mass spectrometric detection; monitoring of the residual drugs in biological fluids for evaluating the efficiency of drug therapy; testing of the enantiomeric purity of pharmaceutical products; the use of novel materials as components of stationary and pseudo-stationary phases in capillary electrophoresis and capillary electrochromatography to increase the selectivity of separation of components of complex matrices; and identification of various on-line preconcentration techniques to reduce the detection limits of biologically active analytes. A topical trend in capillary electrophoresis required in clinical practice, viz., the design of microfluidic systems, is discussed. The bibliography includes 173 references.

  2. The ARM Best Estimate 2-dimensional Gridded Surface

    SciTech Connect

    Xie,Shaocheng; Qi, Tang

    2015-06-15

    The ARM Best Estimate 2-dimensional Gridded Surface (ARMBE2DGRID) data set merges together key surface measurements at the Southern Great Plains (SGP) sites and interpolates the data to a regular 2D grid to facilitate data application. Data from the original site locations can be found in the ARM Best Estimate Station-based Surface (ARMBESTNS) data set.

  3. Integration of 2-Dimensional Materials for Thermoelectric Power Generation

    NASA Astrophysics Data System (ADS)

    Alsaffar, Fadhel; Al Hussain, Abdulrahman; Amer, Moh. R.; Center of Exclence for Green Nanotechnologies Collaboration; Department of Electrical Engineering (UCLA) Collaboration

    Recent developments in nanomaterial research have significantly progressed the performance of thermoelectric devices. Theoretical investigations of the thermoelectic properties of 2-Dimentional monolayers demonstrate a high figure of merit (ZT) .. Here, we investigate the integration of these 2-Dimensional materials for power generation applications using solar heat. We show that using black phosphorus monolayer (phosphorene) as the p-type material, and Molybdenum disulfide (MoS2) monolayers as the n-type material, we get an effective figure of merit (ZT) at least (1.5) with a conversion efficiency of 13% at 280oC. Our results suggest that the integration of various 2-Dimensional materials is a promising approach for commercial thermoelectric power generation applications.

  4. Preparative electrophoresis of living lymphocytes

    NASA Technical Reports Server (NTRS)

    Vanoss, C. J.; Bigazzi, P. E.; Gillman, C. F.; Allen, R. E.

    1974-01-01

    Vertical liquid columns containing low molecular weight dextran density gradients can be used for preparative lymphocyte electrophoresis on earth, in simulation of 0 gravity conditions. Another method that has been tested at 1 G, is the electrophoresis of lymphocytes in a upward direction in vertical columns. By both methods up to 10 to the 7th power lymphocytes can be separated at one time in a 30 cm glass column of 8 mm inside diameter, at 12 v/cm, in 2 hours. Due to convection and sedimentation problems, the separation at 1 G is less than ideal, but it is expected that at 0 gravity electrophoresis will prove to be a uniquely powerful cell separation tool. The technical feasibility of electrophoresing inert particles at 0 G has been proven earlier, during the flight of Apollo 16.

  5. [Mechanisms of organic drug electrophoresis].

    PubMed

    Nuritdinov, V A

    1990-01-01

    Riboflavin, riboflavin mononucleotide, and fluorescein penetration from the ocular bath without electric current (control), from the anode, from the cathode, and without current after preliminary galvanization was studied in rabbit eyes. Penetration of organic drugs into intraocular fluid in electrophoresis was found to be increased not only from the adequate pole of the active electrode, but from the contrary pole as well. Galvanic current sharply abated the barrier function of the cornea. Therapeutic electrophoresis of the eyeball permits the use of many-component solutions with drug ions of different charge. PMID:2238327

  6. Fluid flow electrophoresis in space

    NASA Technical Reports Server (NTRS)

    Griffin, R. N.

    1975-01-01

    Four areas relating to free-flow electrophoresis in space were investigated. The first was the degree of improvement over earthbound operations that might be expected. The second area of investigation covered the problems in developing a flowing buffer electrophoresis apparatus. The third area of investigation was the problem of testing on the ground equipment designed for use in space. The fourth area of investigation was the improvement to be expected in space for purification of biologicals. The results of some ground-based experiments are described. Other studies included cooling requirements in space, fluid sealing techniques, and measurement of voltage drop across membranes.

  7. Bioprocessing: Prospects for space electrophoresis

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1977-01-01

    The basic principles of electrophoresis are reviewed in light of its past contributions to biology and medicine. The near-zero gravity environment of orbiting spacecraft may present some unique advantages for a variety of processes, by abolishing the major source of convection in fluids. As the ground-based development of electrophoresis was heavily influenced by the need to circumvent the effects of gravity, this process should be a prime candidate for space operation. Nevertheless, while a space facility for electrophoresis may overcome the limitations imposed by gravity, it will not necessarily overcome all problems inherent in electrophoresis. These are, mainly, electroosmosis and the dissipation of the heat generated by the electric field. The NASA program has already led to excellent coatings to prevent electroosmosis, while the need for heat dissipation will continue to impose limits on the actual size of equipment. It is also not excluded that, once the dominant force of gravity is eliminated, disturbances in fluid stability may originate from weaker forces, such as surface tension.

  8. DNA electrophoresis in microfabricated devices

    NASA Astrophysics Data System (ADS)

    Dorfman, Kevin D.

    2010-10-01

    Picking up at the conclusion of Viovy’s review of the physics of gel electrophoresis [J.-L. Viovy, Rev. Mod. Phys. 72, 813 (2000)10.1103/RevModPhys.72.813], this review synthesizes the experimental data, theoretical models, and simulation results for DNA electrophoresis in microfabricated and nanofabricated devices appearing since the seminal paper by Volkmuth and Austin [Nature (London) 358, 600 (1992)10.1038/358600a0]. Prototype versions of these devices separate DNA by molecular weight at a rate far superior to gel electrophoresis. After providing an overview of the requisite background material in polymer physics, electrophoresis, and microfluidic device fabrication, the focus is on the following three generic problems: (i) collision with an isolated post, (ii) transport in an array of posts, and (iii) entropic trapping and filtration in the slit-well motif. The transport phenomena are examined here in the context of the length and time scales characterizing the DNA, the device, and the applied electric field.

  9. A tunable isoelectric focusing via moving reaction boundary for two-dimensional gel electrophoresis and proteomics.

    PubMed

    Guo, Chen-Gang; Shang, Zhi; Yan, Jian; Li, Si; Li, Guo-Qing; Liu, Rong-Zhong; Qing, Ying; Fan, Liu-Yin; Xiao, Hua; Cao, Cheng-Xi

    2015-05-01

    Routine native immobilized pH gradient isoelectric focusing (IPG-IEF) and two-dimensional gel electrophoresis (2DE) are still suffering from unfortunate reproducibility, poor resolution (caused by protein precipitation) and instability in characterization of intact protein isoforms and posttranslational modifications. Based on the concept of moving reaction boundary (MRB), we firstly proposed a tunable non-IPG-IEF system to address these issues. By choosing proper pairs of catholyte and anolyte, we could achieve desired cathodic and anodic migrating pH gradients in non-IPG-IEF system, effectively eliminating protein precipitation and uncertainty of quantitation existing in routine IEF and 2DE, and enhancing the resolution and sensitivity of IEF. Then, an adjustable 2DE system was developed by combining non-IPG-IEF with polyacrylamide gel electrophoresis (PAGE). The improved 2DE was evaluated by testing model proteins and colon cancer cell lysates. The experiments revealed that (i) a tunable pH gradient could be designed via MRB; (ii) up to 1.65 fold improvement of resolution was achieved via non-IPG-IEF; (iii) the sensitivity of developed techniques was increased up to 2.7 folds; and (iv) up to about 16.4% more protein spots could be observed via the adjustable 2DE as compared with routine one. The developed techniques might contribute to complex proteome research, especially for screening of biological marker and analysis of extreme acidic/alkaline proteins. PMID:25770625

  10. Techniques For Focusing In Zone Electrophoresis

    NASA Technical Reports Server (NTRS)

    Sharnez, Rizwan; Twitty, Garland E.; Sammons, David W.

    1994-01-01

    In two techniques for focusing in zone electrophoresis, force of applied electrical field in each charged particle balanced by restoring force of electro-osmosis. Two techniques: velocity-gradient focusing (VGF), suitable for rectangular electrophoresis chambers; and field-gradient focusing (FGF), suitable for step-shaped electrophoresis chambers.

  11. Two-Dimensional Gel Electrophoresis: Discovering Biomolecules for Environmental Bioremediation

    NASA Astrophysics Data System (ADS)

    Singh, Om V.; Chandel, Anuj K.

    Environmental contamination has been viewed as an ecological malaise for which bioremediation can be prescribed as a “perfect medicine.” The solution to the problems with bioremediation lies in analyzing to what extent the microbes’ physiological machinery contributes to the degradation process and which biomolecules and their mechanisms are responsible for regulatory factors within the degradation system, such as protein, metabolite, and enzymatic chemical transformation. In the post-genomic era, recent advances in proteomics have allowed us to elucidate many complex biological mechanisms. Two-dimensional gel electrophoresis (2DE) in conjunction with mass spectrometry (MS) can be utilized to identify the biomolecules and their molecular mechanisms in bioremediation. A set of highly abundant global proteins over a pI range 4-7 was separated and compared by size fractionation (25-100 kDa) on 2DE. We identified a set of catabolic proteins, enzymes, and heat shock molecular chaperones associated with the regulatory network that was found to be overexpressed under phenol-stressed conditions. This chapter also offers optimized ideal directions for 2DE, followed by easy-to-follow directions for a protein identification strategy using MALDI-TOF and targeting novel proteins/enzymes for a universal set of experiments.

  12. Electrophoresis technology experiment MA-011

    NASA Technical Reports Server (NTRS)

    Allen, R. E.; Barlow, G. H.; Bier, M.; Bigazzi, P. E.; Knox, R. J.; Micale, F. J.; Seaman, G. V. F.; Vanderhoff, J. W.; Vanoss, C. J.; Patterson, W. J.

    1976-01-01

    Experiment MA-011, electrophoresis technology, was designed to test electrophoresis hardware that would continue the development of technology for electrophoretic separation of materials in the near zero g environment of space. The experimental hardware generally functioned as planned. Frozen live cells were successfully transported into space, electrophoretic processing was performed, and viable cells were returned to earth. A separation of the three types of fixed red blood cells (rabbit, human, and horse) was demonstrated. The human lymphocytes, however, showed no apparent migration. The separation of human kidney cells produced the most exciting data. Analysis shows electrophoretic separation throughout the entire column with at least four bands of viable cells. The isotachophoresis experiment definitely demonstrated the isotachophoretic separation of biological cells in a near zero g environment.

  13. Ratcheted electrophoresis of Brownian particles

    NASA Astrophysics Data System (ADS)

    Kowalik, Mikołaj; Bishop, Kyle J. M.

    2016-05-01

    The realization of nanoscale machines requires efficient methods by which to rectify unbiased perturbations to perform useful functions in the presence of significant thermal noise. The performance of such Brownian motors often depends sensitively on their operating conditions—in particular, on the relative rates of diffusive and deterministic motions. In this letter, we present a type of Brownian motor that uses contact charge electrophoresis of a colloidal particle within a ratcheted channel to achieve directed transport or perform useful work against an applied load. We analyze the stochastic dynamics of this model ratchet to show that it functions under any operating condition—even in the limit of strong thermal noise and in contrast to existing ratchets. The theoretical results presented here suggest that ratcheted electrophoresis could provide a basis for electrochemically powered, nanoscale machines capable of transport and actuation of nanoscale components.

  14. Electrophoresis in strong electric fields.

    PubMed

    Barany, Sandor

    2009-01-01

    Two kinds of non-linear electrophoresis (ef) that can be detected in strong electric fields (several hundred V/cm) are considered. The first ("classical" non-linear ef) is due to the interaction of the outer field with field-induced ionic charges in the electric double layer (EDL) under conditions, when field-induced variations of electrolyte concentration remain to be small comparatively to its equilibrium value. According to the Shilov theory, the non-linear component of the electrophoretic velocity for dielectric particles is proportional to the cubic power of the applied field strength (cubic electrophoresis) and to the second power of the particles radius; it is independent of the zeta-potential but is determined by the surface conductivity of particles. The second one, the so-called "superfast electrophoresis" is connected with the interaction of a strong outer field with a secondary diffuse layer of counterions (space charge) that is induced outside the primary (classical) diffuse EDL by the external field itself because of concentration polarization. The Dukhin-Mishchuk theory of "superfast electrophoresis" predicts quadratic dependence of the electrophoretic velocity of unipolar (ionically or electronically) conducting particles on the external field gradient and linear dependence on the particle's size in strong electric fields. These are in sharp contrast to the laws of classical electrophoresis (no dependence of V(ef) on the particle's size and linear dependence on the electric field gradient). A new method to measure the ef velocity of particles in strong electric fields is developed that is based on separation of the effects of sedimentation and electrophoresis using videoimaging and a new flowcell and use of short electric pulses. To test the "classical" non-linear electrophoresis, we have measured the ef velocity of non-conducting polystyrene, aluminium-oxide and (semiconductor) graphite particles as well as Saccharomice cerevisiae yeast cells as a

  15. Capillary electrophoresis systems and methods

    DOEpatents

    Dorairaj, Rathissh; Keynton, Robert S.; Roussel, Thomas J.; Crain, Mark M.; Jackson, Douglas J.; Walsh, Kevin M.; Naber, John F.; Baldwin, Richard P.; Franco, Danielle B.

    2011-08-02

    An embodiment of the invention is directed to a capillary electrophoresis apparatus comprising a plurality of separation micro-channels. A sample loading channel communicates with each of the plurality of separation channels. A driver circuit comprising a plurality of electrodes is configured to induce an electric field across each of the plurality of separation channels sufficient to cause analytes in the samples to migrate along each of the channels. The system further comprises a plurality of detectors configured to detect the analytes.

  16. Enhancing Centrifugal Separation With Electrophoresis

    NASA Technical Reports Server (NTRS)

    Herrmann, F. T.

    1986-01-01

    Separation of biological cells by coil-planet centrifuge enhanced by electrophoresis. By itself, coil-planet centrifuge offers relatively gentle method of separating cells under low centrifugal force in physiological medium that keeps cells alive. With addition of voltage gradient to separation column of centrifuge, separation still gentle but faster and more complete. Since separation apparatus contains no rotary seal, probability of leakage, contamination, corrosion, and short circuits reduced.

  17. DNA Electrophoresis on Nanopatterned surfaces

    NASA Astrophysics Data System (ADS)

    Luo, Haobin; Gersappe, Dilip

    2002-03-01

    Conventional electrophoretic methods for DNA separation use topological restriction such as the network point in a gel to separate DNA. Here, we present a new approach to DNA electrophoresis using a nanopatterned surface. We exploit the conformational differences that arise when chains of different length are adsorbed onto a flat nanopatterned surface. We use a MD simulation to determine conditions that will optimize separation and present guidelines for the design of nanopatterned surfaces that can separate DNA in a broad size range.

  18. Capillary Electrophoresis - Optical Detection Systems

    SciTech Connect

    Sepaniak, M. J.

    2001-08-06

    Molecular recognition systems are developed via molecular modeling and synthesis to enhance separation performance in capillary electrophoresis and optical detection methods for capillary electrophoresis. The underpinning theme of our work is the rational design and development of molecular recognition systems in chemical separations and analysis. There have been, however, some subtle and exciting shifts in our research paradigm during this period. Specifically, we have moved from mostly separations research to a good balance between separations and spectroscopic detection for separations. This shift is based on our perception that the pressing research challenges and needs in capillary electrophoresis and electrokinetic chromatography relate to the persistent detection and flow rate reproducibility limitations of these techniques (see page 1 of the accompanying Renewal Application for further discussion). In most of our work molecular recognition reagents are employed to provide selectivity and enhance performance. Also, an emerging trend is the use of these reagents with specially-prepared nano-scale materials. Although not part of our DOE BES-supported work, the modeling and synthesis of new receptors has indirectly supported the development of novel microcantilevers-based MEMS for the sensing of vapor and liquid phase analytes. This fortuitous overlap is briefly covered in this report. Several of the more significant publications that have resulted from our work are appended. To facilitate brevity we refer to these publications liberally in this progress report. Reference is also made to very recent work in the Background and Preliminary Studies Section of the Renewal Application.

  19. DNA Sequencing by Capillary Electrophoresis

    PubMed Central

    Karger, Barry L.; Guttman, Andras

    2009-01-01

    Sequencing of human and other genomes has been at the center of interest in the biomedical field over the past several decades and is now leading toward an era of personalized medicine. During this time, DNA sequencing methods have evolved from the labor intensive slab gel electrophoresis, through automated multicapillary electrophoresis systems using fluorophore labeling with multispectral imaging, to the “next generation” technologies of cyclic array, hybridization based, nanopore and single molecule sequencing. Deciphering the genetic blueprint and follow-up confirmatory sequencing of Homo sapiens and other genomes was only possible by the advent of modern sequencing technologies that was a result of step by step advances with a contribution of academics, medical personnel and instrument companies. While next generation sequencing is moving ahead at break-neck speed, the multicapillary electrophoretic systems played an essential role in the sequencing of the Human Genome, the foundation of the field of genomics. In this prospective, we wish to overview the role of capillary electrophoresis in DNA sequencing based in part of several of our articles in this journal. PMID:19517496

  20. Finite temperature holographic duals of 2-dimensional BCFTs

    NASA Astrophysics Data System (ADS)

    Estes, J.

    2015-07-01

    We consider holographic duals of 2-dimensional conformal field theories in the presence of a boundary, interface, defect and/or junction, referred to collectively as BCFTs. In general, the presence of a boundary reduces the SO(2, 2) conformal symmetry to SO(2, 1) and the dual geometry is realized as a warped product of the form , where is not compact. In particular, it will contain points where the warp factor of the AdS 2 space diverges, leading to asymptotically AdS 3 regions. We show that the AdS 2 space-time may always be replaced with an AdS 2-"black-hole" space-time. We argue the resulting geometry describes the BCFT at finite temperature. To motivate this claim, we compute the entanglement entropy holographically for a segment centered around the defect or ending on the boundary and find agreement with a known universal formula.

  1. Mathematical models of continuous flow electrophoresis: Electrophoresis technology

    NASA Technical Reports Server (NTRS)

    Saville, Dudley A.

    1986-01-01

    Two aspects of continuous flow electrophoresis were studied: (1) the structure of the flow field in continuous flow devices; and (2) the electrokinetic properties of suspended particles relevant to electrophoretic separations. Mathematical models were developed to describe flow structure and stability, with particular emphasis on effects due to buoyancy. To describe the fractionation of an arbitrary particulate sample by continuous flow electrophoresis, a general mathematical model was constructed. In this model, chamber dimensions, field strength, buffer composition, and other design variables can be altered at will to study their effects on resolution and throughput. All these mathematical models were implemented on a digital computer and the codes are available for general use. Experimental and theoretical work with particulate samples probed how particle mobility is related to buffer composition. It was found that ions on the surface of small particles are mobile, contrary to the widely accepted view. This influences particle mobility and suspension conductivity. A novel technique was used to measure the mobility of particles in concentrated suspensions.

  2. Proteomic Profiling Of Two-Dimensional Gel Electrophoresis Protein Expression Data

    NASA Astrophysics Data System (ADS)

    Ahmad, Norhaiza; Zhang, J.; Brown, P. J.; James, D. C.; Birch, J. R.; Racher, A. J.; Smales, C. M.

    2008-01-01

    We have undertaken two-dimensional gel electrophoresis (2-DE) proteomic profiling on a series of cell lines with different recombinant antibody production rates. Due to the nature of 2-DE proteomic investigations there will always be `process variability' factors in any data set collected in this way. Some of this variation will arise during sample preparation, gel running and staining, while further variation will arise from the gel analysis procedure. Therefore, in order to identify all significant changes in protein expression between biological samples when analysed by 2-DE, the system precision or `error', and how this correlates to protein abundance, must be known. Only then can the system be considered robust and investigators accurately and confidently report all observable statistically significant changes in protein expression. We introduce an expression variability test to identify protein spots whose expression correlates with increased antibody production. The results have highlighted a small number of candidate proteins for further investigation.

  3. Molecular phylogeny of the hominoid primates as indicated by two-dimensional protein electrophoresis

    SciTech Connect

    Goldman, D.; Giri, P.R.; O'Brien, J.O.

    1987-05-01

    A molecular phylogeny for the hominoid primates was constructed by using genetic distances from a survey of 383 radiolabeled fibroblast polypeptides resolved by two-dimensional electrophoresis (2DE). An internally consistent matrix of Nei genetic distances was generated on the basis of variants in electrophoretic position. The derived phylogenetic tree indicated a branching sequence, from oldest to most recent, of cercopithecoids (Macaca fascicularis), gibbon-siamang, orangutan, gorilla, and human-chimpanzee. A cladistic analysis of 240 electrophoretic characters that varied between ape species produced an identical tree. Genetic distance measures obtained by 2DE are largely consistent with those generated by other molecular procedures. In addition, the 2DE data set appears to resolve the human-chimpanzee-gorilla trichotomy in favor of a more recent association of chimpanzees and humans.

  4. Microbeam-coupled capillary electrophoresis.

    PubMed

    Garty, G; Ehsan, M U; Buonanno, M; Yang, Z; Sweedler, J V; Brenner, D J

    2015-09-01

    Within the first few microseconds following a charged particle traversal of a cell, numerous oxygen and nitrogen radicals are formed along the track. Presented here is a method, using capillary electrophoresis, for simultaneous measurement, within an individual cell, of specific reactive oxygen species, such as the superoxide radical ([Formula: see text]) as well as the native and oxidised forms of glutathione, an ubiquitous anti-oxidant that assists the cell in coping with these species. Preliminary data are presented as well as plans for integrating this system into the charged particle microbeam at Columbia University. PMID:25870435

  5. Integrated multiplexed capillary electrophoresis system

    SciTech Connect

    Yeung, Edward S.; Tan, Hongdong

    2002-05-14

    The present invention provides an integrated multiplexed capillary electrophoresis system for the analysis of sample analytes. The system integrates and automates multiple components, such as chromatographic columns and separation capillaries, and further provides a detector for the detection of analytes eluting from the separation capillaries. The system employs multiplexed freeze/thaw valves to manage fluid flow and sample movement. The system is computer controlled and is capable of processing samples through reaction, purification, denaturation, pre-concentration, injection, separation and detection in parallel fashion. Methods employing the system of the invention are also provided.

  6. Capillary electrophoresis in metallodrug development.

    PubMed

    Holtkamp, Hannah; Hartinger, Christian G

    2015-09-01

    Capillary electrophoresis (CE) is a separation method based on differential migration of analytes in electric fields. The compatibility with purely aqueous separation media makes it a versatile tool in metallodrug research. Many metallodrugs undergo ligand exchange reactions that can easily be followed with this method and the information gained can even be improved by coupling the CE to advanced detectors, such as mass spectrometers. This gives the method high potential to facilitate the development of metallodrugs, especially when combined with innovative method development and experimental design. PMID:26547417

  7. Capillary electrophoresis for drug analysis

    NASA Astrophysics Data System (ADS)

    Lurie, Ira S.

    1999-02-01

    Capillary electrophoresis (CE) is a high resolution separation technique which is amenable to a wide variety of solutes, including compounds which are thermally degradable, non-volatile and highly polar, and is therefore well suited for drug analysis. Techniques which have been used in our laboratory include electrokinetic chromatography (ECC), free zone electrophoresis (CZE) and capillary electrochromatography (CEC). ECC, which uses a charged run buffer additive which migrates counter to osmotic flow, is excellent for many applications, including, drug screening and analyses of heroin, cocaine and methamphetamine samples. ECC approaches include the use of micelles and charged cyclodextrins, which allow for the separation of complex mixtures. Simultaneous separation of acidic, neutral and basic solutes and the resolution of optical isomers and positional isomers are possible. CZE has been used for the analysis of small ions (cations and anions) in heroin exhibits. For the ECC and CZE experiments performed in our laboratory, uncoated capillaries were used. In contrast, CEC uses capillaries packed with high performance liquid chromatography stationary phases, and offers both high peak capacities and unique selectivities. Applications include the analysis of cannabinoids and drug screening. Although CE suffers from limited concentration sensitivity, it is still applicable to trace analysis of drug samples, especially when using injection techniques such as stacking, or detection schemes such as laser induced fluorescence and extended pathlength UV.

  8. Conducting Polymer Electrodes for Gel Electrophoresis

    PubMed Central

    Bengtsson, Katarina; Nilsson, Sara; Robinson, Nathaniel D.

    2014-01-01

    In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that π-conjugated polymers such as poly(3,4-ethylenedioxythiophene) (PEDOT) can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis) systems. In this report, we extend our previous result to gel electrophoresis, and show that electrodes containing PEDOT can be used with a commercial polyacrylamide gel electrophoresis system with minimal impact to the resulting gel image or the ionic transport measured during a separation. PMID:24586761

  9. Electrophoresis-mass spectrometry probe

    DOEpatents

    Andresen, B.D.; Fought, E.R.

    1987-11-10

    The invention involves a new technique for the separation of complex mixtures of chemicals, which utilizes a unique interface probe for conventional mass spectrometers which allows the electrophoretically separated compounds to be analyzed in real-time by a mass spectrometer. This new chemical analysis interface, which couples electrophoresis with mass spectrometry, allows complex mixtures to be analyzed very rapidly, with much greater specificity, and with greater sensitivity. The interface or probe provides a means whereby large and/or polar molecules in complex mixtures to be completely characterized. The preferred embodiment of the probe utilizes a double capillary tip which allows the probe tip to be continually wetted by the buffer, which provides for increased heat dissipation, and results in a continually operating interface which is more durable and electronically stable than the illustrated single capillary tip probe interface. 8 figs.

  10. Electrophoresis-mass spectrometry probe

    DOEpatents

    Andresen, Brian D.; Fought, Eric R.

    1987-01-01

    The invention involves a new technique for the separation of complex mixtures of chemicals, which utilizes a unique interface probe for conventional mass spectrometers which allows the electrophoretically separated compounds to be analyzed in real-time by a mass spectrometer. This new chemical analysis interface, which couples electrophoresis with mass spectrometry, allows complex mixtures to be analyzed very rapidly, with much greater specificity, and with greater sensitivity. The interface or probe provides a means whereby large and/or polar molecules in complex mixtures to be completely characterized. The preferred embodiment of the probe utilizes a double capillary tip which allows the probe tip to be continually wetted by the buffer, which provides for increased heat dissipation, and results in a continually operating interface which is more durable and electronically stable than the illustrated single capillary tip probe interface.

  11. Non-Aqueous Capillary Electrophoresis

    NASA Astrophysics Data System (ADS)

    Szumski, Michał; Buszewski, Bogusław

    Non-aqueous capillary electrophoresis and capillary electrochromatography are special variants of these techniques. Here, organic solvents or their mixtures with or without dissolved electrolytes are used as separation buffer or mobile phase, respectively. The most important features of non-aqueous systems are: better solubility of more hydrophobic ionic substances (many natural products) than in water, much less current and Joule heating allows for using highly concentrated buffers and/or larger capillary internal diameters, polar interactions are enhanced in organic solvents which is often highly advantageous in chiral separation systems. This chapter presents most frequently used solvents, their properties, as well as shows pH* scale which is often used in non-aqueous systems.

  12. Analytical biotechnology: Capillary electrophoresis and chromatography

    SciTech Connect

    Horvath, C.; Nikelly, J.G.

    1990-01-01

    The papers describe the separation, characterization, and equipment required for the electrophoresis or chromatography of cyclic nucleotides, pharmaceuticals, therapeutic proteins, recombinant DNA products, pheromones, peptides, and other biological materials. One paper, On-column radioisotope detection for capillary electrophoresis, has been indexed separately for inclusion on the data base.

  13. Compensating for Electro-Osmosis in Electrophoresis

    NASA Technical Reports Server (NTRS)

    Rhodes, Percy H.; Snyder, Robert S.

    1987-01-01

    Simple mechanical adjustment eliminates transverse velocity component. New apparatus for moving-wall electrophoresis increases degree of collimation of chemical species in sample stream. Electrophoresis chamber set at slight angle in horizontal plane to adjust angle between solution flow and wall motion. Component of velocity created cancels electro-osmotic effect.

  14. Getting the Most out of Electrophoresis Units

    ERIC Educational Resources Information Center

    Mulvihill, Charlotte

    2007-01-01

    At Oklahoma City Community College, they have developed gel electrophoresis activities that support active learning of many scientific concepts, including: pH, electrolysis, oxidation reduction, electrical currents, potentials, conductivity, molarity, gel electrophoresis, DNA and protein separation, and DNA fingerprinting. This article presents…

  15. Fluorescence detection for gel and capillary electrophoresis

    SciTech Connect

    Hogan, B.

    1992-07-21

    First, an indirect fluorescence detection system for the separation of proteins via gel electrophoresis. Quantities as low as 50 nanograms of bovine serum albumin and soybean trypsin inhibitor are separated and detected visually without the need for staining of the analytes. This is very similar to levels of protein commonly separated with gel electrophoresis.

  16. Two Electrophoresis Experiments for Freshmen in the Health Professions.

    ERIC Educational Resources Information Center

    Brabson, G. Dana; Waugh, David S.

    1986-01-01

    Describes procedures involved with paper electrophoresis separation of amino acids, gel electrophoresis separation of DNA, and design of an electrophoresis tank. Describes experiments using paper (amino acids) and gel (deoxyribonucleic acid fragments). Provides material lists, procedures, and discussion. (JM)

  17. Hydrometeor classification from a 2 dimensional video disdrometer

    NASA Astrophysics Data System (ADS)

    Grazioli, Jacopo; Tuia, Devis; Berne, Alexis

    2014-05-01

    Hydrometeor classification techniques aim at identifying the dominant hydrometeor type in a given observation volume or at a given time step, during precipitation. Such techniques are employed to interpret measurements from polarimetric weather radars, cloud lidars, and airborne particle imagers and their output is applied to risk assessment, air traffic control, and parametrization of numerical weather models. In the present work we develop a hydrometeor classification approach designed for data collected by a ground instrument: the 2 dimensional video disdrometer (2DVD). The 2DVD provides fall velocity and 2D views of each particle falling in its sampling area, by means of two orthogonally oriented line scanning cameras. We summarize this large amount of information over time steps of 60 seconds by characterizing the statistical behavior of a set of shape, size and velocity descriptors calculated for each falling hydrometeor. This summarized information is the input for the classification algorithm, that therefore provides the dominant hydrometeor type during a given time step of precipitation. 8 dominant hydrometeor classes have been identified by visual inspection of data collected in different climatologies (Switzerland, France and Canada), namely: small particles, dendrites, columns, graupel, rimed particles, aggregates, melting snow and rain. 400 representative time steps have been manually selected and classified in one of these classes in order to build a training set for the classification algorithm. The employed classifier is a support vector machine (SVM), a supervised linear classification method trained and evaluated on subsets of the 400 time steps. The algorithm achieves accurate performances, with overall accuracy higher than 90% in global terms and higher than 84% in median for each of the 8 hydrometeor classes available. This is confirmed by the Cohen's Kappa score (or HSS), that takes into account the prediction by chance and is higher than 0

  18. Materials Science and Device Physics of 2-Dimensional Semiconductors

    NASA Astrophysics Data System (ADS)

    Fang, Hui

    Materials and device innovations are the keys to future technology revolution. For MOSFET scaling in particular, semiconductors with ultra-thin thickness on insulator platform is currently of great interest, due to the potential of integrating excellent channel materials with the industrially mature Si processing. Meanwhile, ultra-thin thickness also induces strong quantum confinement which in turn affect most of the material properties of these 2-dimensional (2-D) semiconductors, providing unprecedented opportunities for emerging technologies. In this thesis, multiple novel 2-D material systems are explored. Chapter one introduces the present challenges faced by MOSFET scaling. Chapter two covers the integration of ultrathin III V membranes with Si. Free standing ultrathin III-V is studied to enable high performance III-V on Si MOSFETs with strain engineering and alloying. Chapter three studies the light absorption in 2-D membranes. Experimental results and theoretical analysis reveal that light absorption in the 2-D quantum membranes is quantized into a fundamental physical constant, where we call it the quantum unit of light absorption, irrelevant of most of the material dependent parameters. Chapter four starts to focus on another 2-D system, atomic thin layered chalcogenides. Single and few layered chalcogenides are first explored as channel materials, with focuses in engineering the contacts for high performance MOSFETs. Contact treatment by molecular doping methods reveals that many layered chalcogenides other than MoS2 exhibit good transport properties at single layer limit. Finally, Chapter five investigated 2-D van der Waals heterostructures built from different single layer chalcogenides. The investigation in a WSe2/MoS2 hetero-bilayer shows a large Stokes like shift between photoluminescence peak and lowest absorption peak, as well as strong photoluminescence intensity, consistent with spatially indirect transition in a type II band alignment in this

  19. DNA Sequencing Using capillary Electrophoresis

    SciTech Connect

    Dr. Barry Karger

    2011-05-09

    The overall goal of this program was to develop capillary electrophoresis as the tool to be used to sequence for the first time the Human Genome. Our program was part of the Human Genome Project. In this work, we were highly successful and the replaceable polymer we developed, linear polyacrylamide, was used by the DOE sequencing lab in California to sequence a significant portion of the human genome using the MegaBase multiple capillary array electrophoresis instrument. In this final report, we summarize our efforts and success. We began our work by separating by capillary electrophoresis double strand oligonucleotides using cross-linked polyacrylamide gels in fused silica capillaries. This work showed the potential of the methodology. However, preparation of such cross-linked gel capillaries was difficult with poor reproducibility, and even more important, the columns were not very stable. We improved stability by using non-cross linked linear polyacrylamide. Here, the entangled linear chains could move when osmotic pressure (e.g. sample injection) was imposed on the polymer matrix. This relaxation of the polymer dissipated the stress in the column. Our next advance was to use significantly lower concentrations of the linear polyacrylamide that the polymer could be automatically blown out after each run and replaced with fresh linear polymer solution. In this way, a new column was available for each analytical run. Finally, while testing many linear polymers, we selected linear polyacrylamide as the best matrix as it was the most hydrophilic polymer available. Under our DOE program, we demonstrated initially the success of the linear polyacrylamide to separate double strand DNA. We note that the method is used even today to assay purity of double stranded DNA fragments. Our focus, of course, was on the separation of single stranded DNA for sequencing purposes. In one paper, we demonstrated the success of our approach in sequencing up to 500 bases. Other

  20. Comparison of non-electrophoresis grade with electrophoresis grade BIS in NIPAM polymer gel preparation

    PubMed Central

    Khodadadi, Roghayeh; Khajeali, Azim; Farajollahi, Ali Reza; Hajalioghli, Parisa; Raeisi, Noorallah

    2015-01-01

    Introduction:The main objective of this study was to investigate the possibility of replacing electrophoresis cross-linker with non-electrophoresis N, N′-methylenebisacrylamide (BIS) in N-isopropyl acrylamide (NIPAM) polymer gel and its possible effect on dose response. Methods: NIPAM polymer gel was prepared from non-electrophoresis grade BIS and the relaxation rate (R2) was measured by MR imaging after exposing the gel to gamma radiation from Co-60 source. To compare the response of this gel with the one that contains electrophoresis grade BIS, two sets of NIPAM gel were prepared using electrophoresis and non-electrophoresis BIS and irradiated to different gamma doses. Results: It was found that the dose–response of NIPAM gel made from the non-electrophoresis grade BIS is coincident with that of electrophoresis grade BIS. Conclusion:Taken all, it can be concluded that the non-electrophoresis grade BIS not only is a suitable alternative for the electrophoresis grade BIS but also reduces the cost of gel due to its lower price. PMID:26457250

  1. Nonlinear waves in capillary electrophoresis

    PubMed Central

    Ghosal, Sandip; Chen, Zhen

    2011-01-01

    Electrophoretic separation of a mixture of chemical species is a fundamental technique of great usefulness in biology, health care and forensics. In capillary electrophoresis the sample migrates in a microcapillary in the presence of a background electrolyte. When the ionic concentration of the sample is sufficiently high, the signal is known to exhibit features reminiscent of nonlinear waves including sharp concentration ‘shocks’. In this paper we consider a simplified model consisting of a single sample ion and a background electrolyte consisting of a single co-ion and a counterion in the absence of any processes that might change the ionization states of the constituents. If the ionic diffusivities are assumed to be the same for all constituents the concentration of sample ion is shown to obey a one dimensional advection diffusion equation with a concentration dependent advection velocity. If the analyte concentration is sufficiently low in a suitable non-dimensional sense, Burgers’ equation is recovered, and thus, the time dependent problem is exactly solvable with arbitrary initial conditions. In the case of small diffusivity either a leading edge or trailing edge shock is formed depending on the electrophoretic mobility of the sample ion relative to the background ions. Analytical formulas are presented for the shape, width and migration velocity of the sample peak and it is shown that axial dispersion at long times may be characterized by an effective diffusivity that is exactly calculated. These results are consistent with known observations from physical and numerical simulation experiments. PMID:20238181

  2. Phenomenology of colloidal crystal electrophoresis

    NASA Astrophysics Data System (ADS)

    Medebach, Martin; Palberg, Thomas

    2003-08-01

    We studied the motion of polycrystalline solids comprising of charged sub-micron latex spheres suspended in deionized water. These were subjected to a low frequency alternating square wave electric field in an optical cell of rectangular cross section. Velocity profiles in X and Y direction were determined by Laser Doppler Velocimetry. The observed complex flow profiles are time dependent due to the combined effects of electro-osmosis, electrophoresis, crystal elasticity, and friction of the crystals at the cell wall. On small time scales elastic deformation occurs. On long time scales channel formation is observed. At intermediate times steady state profiles are dominated by a solid plug of polycrystalline material moving in the cell center. At large field strengths the plug shear melts. Mobilities in the shear molten state are on the order of (6.5±0.5) 10-8 m2 V-1 s-1 and connect continuously with those of the equilibrium fluid. The apparent mobility of the plug is much larger than of the fluid and like the mobility of the fluid decreases with increasing particle number density. We qualitatively attribute the accelerated motion of the plug to an incomplete exposure to the electro-osmotic flow profile.

  3. Atomic Force Controlled Capillary Electrophoresis

    NASA Astrophysics Data System (ADS)

    Lewis, Aaron; Yeshua, Talia; Palchan, Mila; Lovsky, Yulia; Taha, Hesham

    2010-03-01

    Lithography based on scanning probe microscopic techniques has considerable potential for accurate & localized deposition of material on the nanometer scale. Controlled deposition of metallic features with high purity and spatial accuracy is of great interest for circuit edit applications in the semiconductor industry, for plasmonics & nanophotonics and for basic research in surface enhanced Raman scattering & nanobiophysics. Within the context of metal deposition we will review the development of fountain pen nanochemistry and its most recent emulation Atomic Force Controlled Capillary Electrophoresis (ACCE). Using this latter development we will demonstrate achievement of unprecedented control of nanoparticle deposition using a three-electrode geometry. Three electrodes are attached: one on the outside of a metal coated glass probe, one on the inside of a hollow probe in a solution containing Au nanoparticles in the capillary, and a third on the surface where the writing takes place. The three electrodes provide electrical pulses for accurate control of deposition and retraction of the liquid from the surface overcoming the lack of control seen in both dip pen lithography & fountain pen nanochemistry when the tip contacts the surface. With this development, we demonstrate depositing a single 1.3 nm Au nanoparticle onto surfaces such as semiconductors.

  4. High-performance capillary electrophoresis of histones

    SciTech Connect

    Gurley, L.R.; London, J.E.; Valdez, J.G.

    1991-01-01

    A high performance capillary electrophoresis (HPCE) system has been developed for the fractionation of histones. This system involves electroinjection of the sample and electrophoresis in a 0.1M phosphate buffer at pH 2.5 in a 50 {mu}m {times} 35 cm coated capillary. Electrophoresis was accomplished in 9 minutes separating a whole histone preparation into its components in the following order of decreasing mobility; (MHP) H3, H1 (major variant), H1 (minor variant), (LHP) H3, (MHP) H2A (major variant), (LHP) H2A, H4, H2B, (MHP) H2A (minor variant) where MHP is the more hydrophobic component and LHP is the less hydrophobic component. This order of separation is very different from that found in acid-urea polyacrylamide gel electrophoresis and in reversed-phase HPLC and, thus, brings the histone biochemist a new dimension for the qualitative analysis of histone samples. 27 refs., 8 figs.

  5. Capillary electrophoresis electrospray ionization mass spectrometry interface

    SciTech Connect

    Smith, R.D.; Severs, J.C.

    1999-11-30

    The present invention is an interface between a capillary electrophoresis separation capillary end and an electrospray ionization mass spectrometry emitter capillary end, for transporting an analyte sample from a capillary electrophoresis separation capillary to a electrospray ionization mass spectrometry emitter capillary. The interface of the present invention has: (a) a charge transfer fitting enclosing both of the capillary electrophoresis capillary end and the electrospray ionization mass spectrometry emitter capillary end; (b) a reservoir containing an electrolyte surrounding the charge transfer fitting; and (c) an electrode immersed into the electrolyte, the electrode closing a capillary electrophoresis circuit and providing charge transfer across the charge transfer fitting while avoiding substantial bulk fluid transfer across the charge transfer fitting. Advantages of the present invention have been demonstrated as effective in providing high sensitivity and efficient analyses.

  6. Capillary electrophoresis electrospray ionization mass spectrometry interface

    DOEpatents

    Smith, Richard D.; Severs, Joanne C.

    1999-01-01

    The present invention is an interface between a capillary electrophoresis separation capillary end and an electrospray ionization mass spectrometry emitter capillary end, for transporting an anolyte sample from a capillary electrophoresis separation capillary to a electrospray ionization mass spectrometry emitter capillary. The interface of the present invention has: (a) a charge transfer fitting enclosing both of the capillary electrophoresis capillary end and the electrospray ionization mass spectrometry emitter capillary end; (b) a reservoir containing an electrolyte surrounding the charge transfer fitting; and (c) an electrode immersed into the electrolyte, the electrode closing a capillary electrophoresis circuit and providing charge transfer across the charge transfer fitting while avoiding substantial bulk fluid transfer across the charge transfer fitting. Advantages of the present invention have been demonstrated as effective in providing high sensitivity and efficient analyses.

  7. Maize MeJA-responsive proteins identified by high-resolution 2-DE PAGE.

    PubMed

    Zhang, Yuliang; Pennerman, Kayla K; Yang, Fengshan; Yin, Guohua

    2015-12-01

    Exogenous methyl jasmonate (MeJA) is well-known to induce plant defense mechanisms effective against a wide variety of insect and microbial pests. High-resolution 2-DE gel electrophoresis was used to discover changes in the leaf proteome of maize exposed to MeJA. We sequenced 62 MeJA-responsive proteins by tandem mass spectroscopy, and deposited the mass spectra and identities in the EMBL-EBI PRIDE repository under reference number PXD001793. An analysis and discussion of the identified proteins in relation to maize defense against Asian corn borer is published by Zhang et al. (2015) [1]. PMID:26509185

  8. Microfluidic Breadboard Approach to Capillary Electrophoresis.

    PubMed

    Koenka, Israel Joel; Sáiz, Jorge; Rempel, Paul; Hauser, Peter C

    2016-04-01

    A breadboard approach for electrophoretic separations with contactless conductivity detection is presented. This is based on miniature off-the-shelf components such as syringe pumps, valves, and pressure controllers which could be set up in a very compact overall arrangement. It has a high flexibility for different tasks at hand, and the common operations of hydrodynamic injection and capillary flushing are automated. For demonstration of the versatility of the proposition, several very diverse configurations and modes of electrophoresis were successfully implemented, namely, standard capillary zone electrophoresis, pressure assisted zone electrophoresis, the simultaneous separation of cations and anions by dual-capillary zone electrophoresis, the separation of cationic amino acids by isotachophoresis, as well as the separation of small carboxylic acids by gradient elution moving boundary electrophoresis. The system also allows fast separations, as demonstrated by the analysis of six inorganic cations within 35 s. The approach addresses respective limitations of either conventional capillary electrophoresis instruments as well as electrophoretic lab-on-chip devices, while maintaining a performance in terms of detection limits and reproducibility comparable to standard instrumentation. PMID:26926522

  9. Free flow cell electrophoresis using zwitterionic buffer

    NASA Technical Reports Server (NTRS)

    Rodkey, R. Scott

    1990-01-01

    Studies of a zwitterionic buffer formulated for cell electrophoresis were done using the McDonnell-Douglas Continuous Flow Electrophoresis System. Standard buffers were analyzed for their stability in the electrical field and the results showed that both buffers tested were inherently unstable. Further, titration studies showed that the standards buffers buffered poorly at the pH employed for electrophoresis. The zwitterionic buffer buffered well at its nominal pH and was shown to be stable in the electrical field. Comparative studies of the buffer with standard cell separation buffers using formalin fixed rabbit and goose red blood cells showed that the zwitterionic buffer gave better resolution of the fixed cells. Studies with viable hybridoma cells showed that buffer Q supported cell viability equal to Hank's Balanced Salt Solution and that hybridoma cells in different stages of the growth cycle demonstrated reproducible differences in electrophoretic mobility.

  10. DNA electrophoresis in Pluronic F127

    NASA Astrophysics Data System (ADS)

    You, Seungyong; van Winkle, David

    2006-03-01

    Electrophoresis involves the separation of bio-molecules in a sieving medium by applying an electric field. DNA molecule fragments are separated in conventional gels and a several models have been successfully applied for understanding the separations. Recently, a pluronic gel was found to be an effective sieving medium for electrophoresis. However, the mobility of DNA in this gel cannot be described by the conventional theories. One reason is that Pluronic F127 is not a crosslinked gel, but a lattice of polymer micelles. The migration of single DNA molecules stained with various dye molecules was studied in slab gel electrophoresis by real-time fluorescence microscopy. Results for a variety of sizes will be presented.

  11. Application of Microchip Electrophoresis for Clinical Tests

    NASA Astrophysics Data System (ADS)

    Yatsushiro, Shouki; Kataoka, Masatoshi

    Microchip electrophoresis has recently attracted much attention in the field of nuclear acid analysis due to its high efficiency, ease of operation, low consumption of samples and reagents, and relatively low costs. In addition, the analysis has expanded to an analytical field like not only the analysis of DNA but also the analysis of RNA, the protein, the sugar chain, and the cellular function, etc. In this report, we showed that high-performance monitoring systems for human blood glucose levels and α-amylase activity in human plasma using microchip electrophoresis.

  12. Role of gravity in preparative electrophoresis

    NASA Technical Reports Server (NTRS)

    Bier, M.; Hinckley, J. O. N.; Smolka, A. J. K.; Binder, M. J.; Coxon, M.; Nee, T. W.; Scully, M. O.; Shih, H. S. T.; Snyder, R. S.

    1974-01-01

    Electrophoresis has contributed significantly to the methodology of biological sciences, and shows the potential for large scale fractionation of a wide range of medically important substances, including living cells. Gravity plays an important role in the electrophoretic process, and hence the importance of the NASA effort to develop a zero-gravity separation facility as part of its shuttle program. The current state of art in electrophoresis is reviewed with particular emphasis on the role of gravity and the possible use of istachophoresis. This technique utilizes a discontinuous buffer system, and appears to be the only high resolution electrophoretic technique currently available for separation of living cells.

  13. Undergraduate physics laboratory: Electrophoresis in chromatography paper

    NASA Astrophysics Data System (ADS)

    Hyde, Alexander; Batishchev, Oleg

    2015-12-01

    An experiment studying the physical principles of electrophoresis in liquids was developed for an undergraduate laboratory. We have improved upon the standard agarose gel electrophoresis experimental regime with a straightforward and cost-effective procedure, in which drops of widely available black food coloring were separated by electric field into their dye components on strips of chromatography paper soaked in a baking soda/water solution. Terminal velocities of seven student-safe dyes were measured as a function of the electric potential applied along the strips. The molecular mobility was introduced and calculated by analyzing data for a single dye. Sources of systematic and random errors were investigated.

  14. Two-dimensional electrophoresis analysis of proteins extracted from Alexandrium sp. LC3

    NASA Astrophysics Data System (ADS)

    Li, Hao; Miao, Jinlai; Cui, Fengxia; Li, Guangyou

    2007-10-01

    Two-dimensional electrophoresis(2-DE) of protein extracted and purified from Alexandrium sp. LC3 was conducted. In the SDS-PAGE study, the relative molecular weights of the proteins were mainly in the range of 14kDa-31kDa and 43kDa-66kDa, and more proteins were detected between 14kDa and 31kDa. With the improved protein preparation, the two-dimensional electrophoresis patterns indicated that the relative molecular weights of the proteins were between 14kDa and 100kDa, and most of them ranged from 14kDa to 31kDa. This was consistent with the result of the SDS-PAGE analysis. The isoelectric points were found to lie between 3.0 and 8.0, and most of them were in the range of 3.0 6.0. Better separation effect was acquired with pre-prepared immobilized gradient (IPG) strip (pH3 5.6), and about 320 protein spots could be visualized on the 2-DE map by staining. Within pH3 10 and pH3 5.6 strips, the protein samples of Alexandrium sp. LC3 could be separated well.

  15. DNA ADDUCT RESEARCH WITH CAPILLARY ELECTROPHORESIS

    EPA Science Inventory

    DNA's central importance in all biological systems dictates a wide variety of DNA-related research. or much of this research, the utilization of capillary electrophoresis (CE) can be of significant advantage. pen-tube CE yields excellent separations of DNA components, which can b...

  16. A Simple Vertical Slab Gel Electrophoresis Apparatus.

    ERIC Educational Resources Information Center

    Carter, J. B.; And Others

    1983-01-01

    Describes an inexpensive, easily constructed, and safe vertical slab gel kit used routinely for sodium dodecyl sulphate-polyacrylamide gel electrophoresis research and student experiments. Five kits are run from a single transformer. Because toxic solutions are used, students are given plastic gloves and closely supervised during laboratory…

  17. DNA DAMAGE QUANTITATION BY ALKALINE GEL ELECTROPHORESIS.

    SciTech Connect

    SUTHERLAND,B.M.; BENNETT,P.V.; SUTHERLAND, J.C.

    2004-03-24

    Physical and chemical agents in the environment, those used in clinical applications, or encountered during recreational exposures to sunlight, induce damages in DNA. Understanding the biological impact of these agents requires quantitation of the levels of such damages in laboratory test systems as well as in field or clinical samples. Alkaline gel electrophoresis provides a sensitive (down to {approx} a few lesions/5Mb), rapid method of direct quantitation of a wide variety of DNA damages in nanogram quantities of non-radioactive DNAs from laboratory, field, or clinical specimens, including higher plants and animals. This method stems from velocity sedimentation studies of DNA populations, and from the simple methods of agarose gel electrophoresis. Our laboratories have developed quantitative agarose gel methods, analytical descriptions of DNA migration during electrophoresis on agarose gels (1-6), and electronic imaging for accurate determinations of DNA mass (7-9). Although all these components improve sensitivity and throughput of large numbers of samples (7,8,10), a simple version using only standard molecular biology equipment allows routine analysis of DNA damages at moderate frequencies. We present here a description of the methods, as well as a brief description of the underlying principles, required for a simplified approach to quantitation of DNA damages by alkaline gel electrophoresis.

  18. Planetary In Situ Capillary Electrophoresis System (PISCES)

    NASA Astrophysics Data System (ADS)

    Willis, P. A.; Stockton, A. M.; Mora, M. F.; Cable, M. L.; Bramall, N. E.; Jensen, E. C.; Jiao, H.; Lynch, E.; Mathies, R. A.

    2012-10-01

    We propose to develop PISCES, a 3-kg, 2W, flight-capable microfluidic lab-on-a-chip capillary electrophoresis analyzer capable of ingesting solid, liquid, or gas samples and performing a suite of chemical analyses with parts per trillion sensitivity.

  19. Role of gravity in preparative electrophoresis

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1975-01-01

    The fundamental formulas of electrophoresis are derived microscopically and applied to the problem of isotachophoresis. A simple physical model of the isotachophoresis front is proposed. The front motion and structure are studied in the simplified case without convection, diffusion and non-electric external forces.

  20. Increasing Sensitivity In Continuous-Flow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Sharnez, Rizwan; Sammons, David W.

    1994-01-01

    Sensitivity of continuous-flow electrophoresis (CFE) chamber increased by introducing lateral gradients in concentration of buffer solution and thickness of chamber. Such gradients, with resulting enhanced separation, achieved in CFE chamber with wedge-shaped cross section and collateral flow. Enables improved separations of homogeneous components of mixtures of variety of biologically important substances.

  1. Using Gel Electrophoresis To Illustrate Protein Diversity and Isoelectric Point.

    ERIC Educational Resources Information Center

    Browning, Mark; Vanable, Joseph

    2002-01-01

    Demonstrates the differences in protein structures by focusing on isoelectric point with an experiment that is observable under certain pH levels in gel electrophoresis. Explains the electrophoresis procedure and reports results of the experiments. (YDS)

  2. Statistical analysis on 1-dimensional and 2-dimensional thermal dissipation for nickel metal hydride battery system

    NASA Astrophysics Data System (ADS)

    Hashim, Mohammad Firdaus Abu; Ramakrishnan, Sivakumar; Mohamad, Ahmad Azmin

    2014-06-01

    Due to low environmental impact and rechargeable capability, the Nickel Metal Hydride battery has been considered to be one of the most promising candidate battery for electrical vehicle nowadays. The energy delivered by the Nickel Metal Hydride battery depends heavily on its discharge profile and generally it is intangible to tract the trend of the energydissipation that is stored in the battery for informative analysis. The thermal models were developed in 1-dimensional and 2-dimensional using Matlab and these models are capable of predicting the temperature distributions inside a cell. The simulated results were validated and verified with referred exact sources of experimental data using Minitab software. The result for 1-Dimensional showed that the correlations between experimental and predicted results for the time intervals 60 minutes, 90 minutes, and 114 minutes frompositive to negative electrode thermal dissipationdirection are34%, 83%, and 94% accordingly while for the 2-Dimensional the correlational results for the same above time intervals are44%, 93% and 95%. These correlationalresults between experimental and predicted clearly indicating the thermal behavior under natural convention can be well fitted after around 90 minutes durational time and 2-Dimensional model can predict the results more accurately compared to 1-Dimensional model. Based on the results obtained from simulations, it can be concluded that both 1-Dimensional and 2-Dimensional models can predict nearly similar thermal behavior under natural convention while 2-Dimensional model was used to predict thermal behavior under forced convention for better accuracy.

  3. Characterization of wheat gliadin proteins by combined two-dimensional gel electrophoresis and tandem mass spectrometry.

    PubMed

    Mamone, Gianfranco; Addeo, Francesco; Chianese, Lina; Di Luccia, Aldo; De Martino, Alessandra; Nappo, Annunziata; Formisano, Annarita; De Vivo, Pasqualina; Ferranti, Pasquale

    2005-07-01

    A proteomics-based approach was used for characterizing wheat gliadins from an Italian common wheat (Triticum aestivum) cultivar. A two-dimensional gel electrophoresis (2-DE) map of roughly 40 spots was obtained by submitting the 70% alcohol-soluble crude protein extract to isoelectric focusing on immobilized pH gradient strips across two pH gradient ranges, i.e., 3-10 or pH 6-11, and to sodium dodecyl sulfate-polyacrylamide electrophoresis in the second dimension. The chymotryptic digest of each spot was characterized by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and nano electrospray ionization-tandem mass spectrometry (MS/MS) analysis, providing a "peptide map" for each digest. The measured masses were subsequently sought in databases for sequences. For accurate identification of the parent protein, it was necessary to determine de novo sequences by MS/MS experiments on the peptides. By partial mass fingerprinting, we identified protein molecules such as alpha/beta-, gamma-, omega-gliadin, and high molecular weight-glutenin. The single spots along the 2-DE map were discriminated on the basis of their amino acid sequence traits. alpha-Gliadin, the most represented wheat protein in databases, was highly conserved as the relative N-terminal sequence of the components from the 2-DE map contained only a few silent amino acid substitutions. The other closely related gliadins were identified by sequencing internal peptide chains. The results gave insight into the complex nature of gliadin heterogeneity. This approach has provided us with sound reference data for differentiating gliadins amongst wheat varieties. PMID:15952231

  4. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An electrophoresis... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Electrophoresis apparatus for clinical use....

  5. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An electrophoresis... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Electrophoresis apparatus for clinical use....

  6. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An electrophoresis... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Electrophoresis apparatus for clinical use....

  7. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL CHEMISTRY AND CLINICAL TOXICOLOGY DEVICES Clinical Laboratory Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An electrophoresis... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Electrophoresis apparatus for clinical use....

  8. 21 CFR 862.2485 - Electrophoresis apparatus for clinical use.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Electrophoresis apparatus for clinical use. 862... Instruments § 862.2485 Electrophoresis apparatus for clinical use. (a) Identification. An electrophoresis apparatus for clinical use is a device intended to separate molecules or particles, including...

  9. Proteomic profiling of Plasmodium falciparum through improved, semiquantitative two-dimensional gel electrophoresis.

    PubMed

    Smit, Salome; Stoychev, Stoyan; Louw, Abraham I; Birkholtz, Lyn-Marie

    2010-05-01

    Two-dimensional gel electrophoresis (2-DE) is one of the most commonly used technologies to obtain a snapshot of the proteome at any specific time. However, its application to study the Plasmodial (malaria parasite) proteome is still limited due to inefficient extraction and detection methods and the extraordinarily large size of some proteins. Here, we report an optimized protein extraction method, the most appropriate methods for Plasmodial protein quantification and 2-DE detection, and finally protein identification by mass spectrometry (MS). Linear detection of Plasmodial proteins in a optimized lysis buffer was only possible with the 2-D Quant kit, and of the four stains investigated, Flamingo Pink was superior regarding sensitivity, linearity, and excellent MS-compatibility. 2-DE analyses of the Plasmodial proteome using this methodology resulted in the reliable detection of 349 spots and a 95% success rate in MS/MS identification. Subsequent application to the analyses of the Plasmodial ring and trophozoite proteomes ultimately resulted in the identification of 125 protein spots, which constituted 57 and 49 proteins from the Plasmodial ring and trophozoite stages, respectively. This study additionally highlights the presence of various isoforms within the Plasmodial proteome, which is of significant biological importance within the Plasmodial parasite during development in the intraerythrocytic developmental cycle. PMID:20218691

  10. Analysis of enzyme activity regulation by non-denaturing electrophoresis and application of this regulation for enzyme reactor production.

    PubMed

    Shimazaki, Youji; Miki, Shizuka

    2013-10-01

    Non-denaturing electrophoresis can be used to screen enzymes that self-regulate their activities by using a combination of enzymes and their inhibitors. Furthermore, this technique can be applied to develop enzyme reactors that self-regulate their activities. After separation of proteins from mouse liver cytosol by non-denaturing isoelectric focusing, lactate dehydrogense (LDH) and esterase activities were qualitatively and quantitatively examined using a combination of two-dimensional electrophoresis (2-DE) and non-denaturing stacking gel electrophoresis. Activities of mouse liver-derived LDH and carboxylesterase were reversibly inhibited by oxamate and 6,9-diamino-2-ethoxyacridine (acrinol), respectively, in the stacking gels and recovered when the enzymes migrated towards the separation gels. After separation and immobilization of the enzymes, their activities were inhibited by inhibitors and recovered after inhibitor removal. These results indicate that non-denaturing electrophoresis can be applied to select enzymes that self-regulate their activities and subsequently aid in the development of enzyme reactors that can control the enzyme activities. PMID:22803677

  11. Electrophoresis for genotyping: microtiter array diagonal gel electrophoresis on horizontal polyacrylamide gels, hydrolink, or agarose.

    PubMed

    Day, I N; Humphries, S E

    1994-11-01

    Electrophoresis of DNA has been performed traditionally in either an agarose or acrylamide gel matrix. Considerable effort has been directed to improved quality agaroses capable of high resolution, but for small fragments, such as those from polymerase chain reaction (PCR) and post-PCR digests, acrylamide still offers the highest resolution. Although agarose gels can easily be prepared in an open-faced format to gain the conveniences of horizontal electrophoresis, acrylamide does not polymerize in the presence of air and the usual configurations for gel preparation lead to electrophoresis in the vertical dimension. We describe here a very simple device and method to prepare and manipulate horizontal polyacrylamide gels (H-PAGE). In addition, the open-faced horizontal arrangement enables loading of arrays of wells. Since many procedures are undertaken in standard 96-well microtiter plates, we have also designed a device which preserves the exact configuration of the 8 x 12 array and enables electrophoresis in tracks following a 71.6 degrees diagonal between wells (MADGE, microtiter array diagonal gel electrophoresis), using either acrylamide or agarose. This eliminates almost all of the staff time taken in setup, loading, and recordkeeping and offers high resolution for genotyping pattern recognition. The nature and size of the gels allow direct stacking of gels in one tank, so that a tank used typically to analyze 30-60 samples can readily be used to analyze 1000-2000 samples. The gels would also enable robotic loading. Electrophoresis allows analysis of size and charge, parameters inaccessible to liquid-phase methods: thus, genotyping size patterns, variable length repeats, and haplotypes is possible, as well as adaptability to typing of point variations using protocols which create a difference detectable by electrophoresis. PMID:7864363

  12. Mathematical Models of Continuous Flow Electrophoresis

    NASA Technical Reports Server (NTRS)

    Saville, D. A.; Snyder, R. S.

    1985-01-01

    Development of high resolution continuous flow electrophoresis devices ultimately requires comprehensive understanding of the ways various phenomena and processes facilitate or hinder separation. A comprehensive model of the actual three dimensional flow, temperature and electric fields was developed to provide guidance in the design of electrophoresis chambers for specific tasks and means of interpreting test data on a given chamber. Part of the process of model development includes experimental and theoretical studies of hydrodynamic stability. This is necessary to understand the origin of mixing flows observed with wide gap gravitational effects. To insure that the model accurately reflects the flow field and particle motion requires extensive experimental work. Another part of the investigation is concerned with the behavior of concentrated sample suspensions with regard to sample stream stability particle-particle interactions which might affect separation in an electric field, especially at high field strengths. Mathematical models will be developed and tested to establish the roles of the various interactions.

  13. DNA electrophoresis in a nanofence array†

    PubMed Central

    Park, Sung-Gyu; Olson, Daniel W.

    2012-01-01

    We present the design and implementation of an oxidized silicon “nanofence array” for long DNA electrophoresis. The device consists of a periodic array of post-filled regions (the nanofences) alternating with empty channel regions. Even in this prototype version, the nanofence array provides the resolving power of a hexagonal nanopost array without requiring any direct-write nanopatterning steps such as electron-beam lithography. Through detailed single molecule investigations, we demonstrate that the origin of the resolving power of the nanofence array is not a reduction in band broadening, which might be expected from the theories for DNA electrophoresis in post arrays. Rather, the enhanced stretching of the hooked DNA by the uniform electric field between nanofences increases the efficiency of the collisions. PMID:22388662

  14. Numerical simulation of electrophoresis separation processes

    NASA Technical Reports Server (NTRS)

    Ganjoo, D. K.; Tezduyar, T. E.

    1986-01-01

    A new Petrov-Galerkin finite element formulation has been proposed for transient convection-diffusion problems. Most Petrov-Galerkin formulations take into account the spatial discretization, and the weighting functions so developed give satisfactory solutions for steady state problems. Though these schemes can be used for transient problems, there is scope for improvement. The schemes proposed here, which consider temporal as well as spatial discretization, provide improved solutions. Electrophoresis, which involves the motion of charged entities under the influence of an applied electric field, is governed by equations similiar to those encountered in fluid flow problems, i.e., transient convection-diffusion equations. Test problems are solved in electrophoresis and fluid flow. The results obtained are satisfactory. It is also expected that these schemes, suitably adapted, will improve the numerical solutions of the compressible Euler and the Navier-Stokes equations.

  15. Fish Muscle Proteins: Extraction, Quantitation, and Electrophoresis

    NASA Astrophysics Data System (ADS)

    Smith, Denise

    Electrophoresis can be used to separate and visualize proteins. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), proteins are separated based on size. When protein samples are applied to such gels, it is usually necessary to know the protein content of the sample. This makes it possible to apply a volume of sample to the gel such that samples have a comparable amount of total protein. While it is possible to use an official method of protein analysis (e.g., Kjeldahl, N combustion) for such an application, it often is convenient to use a rapid spectroscopic protein analysis that requires only a small amount of sample. The bicinchoninic acid (BCA) assay method will be used for this purpose.

  16. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, Edward S.; Taylor, John A.

    1994-06-28

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

  17. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, Edward S.; Taylor, John A.

    1996-03-12

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis.

  18. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, E.S.; Taylor, J.A.

    1994-06-28

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis. 14 figures.

  19. Multiplexed fluorescence detector system for capillary electrophoresis

    DOEpatents

    Yeung, E.S.; Taylor, J.A.

    1996-03-12

    A fluorescence detection system for capillary electrophoresis is provided wherein the detection system can simultaneously excite fluorescence and substantially simultaneously monitor separations in multiple capillaries. This multiplexing approach involves laser irradiation of a sample in a plurality of capillaries through optical fibers that are coupled individually with the capillaries. The array is imaged orthogonally through a microscope onto a charge-coupled device camera for signal analysis. 14 figs.

  20. Capillary zone electrophoresis-mass spectrometer interface

    DOEpatents

    D'Silva, Arthur

    1996-08-06

    A device for providing equal electrical potential between two loci unconnected by solid or liquid electrical conducts is provided. The device comprises a first electrical conducting terminal, a second electrical conducting terminal connected to the first terminal by a rigid dielectric structure, and an electrically conducting gas contacting the first and second terminals. This device is particularly suitable for application in the electrospray ionization interface between a capillary zone electrophoresis apparatus and a mass spectrometer.

  1. Capillary zone electrophoresis-mass spectrometer interface

    DOEpatents

    D`Silva, A.

    1996-08-06

    A device for providing equal electrical potential between two loci unconnected by solid or liquid electrical conductors is provided. The device comprises a first electrical conducting terminal, a second electrical conducting terminal connected to the first terminal by a rigid dielectric structure, and an electrically conducting gas contacting the first and second terminals. This device is particularly suitable for application in the electrospray ionization interface between a capillary zone electrophoresis apparatus and a mass spectrometer. 1 fig.

  2. Method and apparatus for continuous electrophoresis

    DOEpatents

    Watson, Jack S.

    1992-01-01

    A method and apparatus for conducting continuous separation of substances by electrophoresis are disclosed. The process involves electrophoretic separation combined with couette flow in a thin volume defined by opposing surfaces. By alternating the polarity of the applied potential and producing reciprocating short rotations of at least one of the surfaces relative to the other, small increments of separation accumulate to cause substantial, useful segregation of electrophoretically separable components in a continuous flow system.

  3. Portable electrophoresis apparatus using minimum electrolyte

    NASA Technical Reports Server (NTRS)

    Stevens, M. R.; Vickers, J. M. (Inventor)

    1976-01-01

    An electrophoresis unit for use in conducting electrophoretic analysis of specimens is described. The unit includes a sealable container in which a substrate mounted specimen is suspended in an electrolytic vapor. A heating unit is employed to heat a supply of electrolyte to produce the vapor. The substrate is suspended within the container by being attached between a pair of clips which also serve as electrodes to which a direct current power source may be connected.

  4. Consensus brain-derived protein, extraction protocol for the study of human and murine brain proteome using both 2D-DIGE and mini 2DE immunoblotting.

    PubMed

    Fernandez-Gomez, Francisco-Jose; Jumeau, Fanny; Derisbourg, Maxime; Burnouf, Sylvie; Tran, Hélène; Eddarkaoui, Sabiha; Obriot, Hélène; Dutoit-Lefevre, Virginie; Deramecourt, Vincent; Mitchell, Valérie; Lefranc, Didier; Hamdane, Malika; Blum, David; Buée, Luc; Buée-Scherrer, Valérie; Sergeant, Nicolas

    2014-01-01

    Two-dimensional gel electrophoresis (2DE) is a powerful tool to uncover proteome modifications potentially related to different physiological or pathological conditions. Basically, this technique is based on the separation of proteins according to their isoelectric point in a first step, and secondly according to their molecular weights by SDS polyacrylamide gel electrophoresis (SDS-PAGE). In this report an optimized sample preparation protocol for little amount of human post-mortem and mouse brain tissue is described. This method enables to perform both two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and mini 2DE immunoblotting. The combination of these approaches allows one to not only find new proteins and/or protein modifications in their expression thanks to its compatibility with mass spectrometry detection, but also a new insight into markers validation. Thus, mini-2DE coupled to western blotting permits to identify and validate post-translational modifications, proteins catabolism and provides a qualitative comparison among different conditions and/or treatments. Herein, we provide a method to study components of protein aggregates found in AD and Lewy body dementia such as the amyloid-beta peptide and the alpha-synuclein. Our method can thus be adapted for the analysis of the proteome and insoluble proteins extract from human brain tissue and mice models too. In parallel, it may provide useful information for the study of molecular and cellular pathways involved in neurodegenerative diseases as well as potential novel biomarkers and therapeutic targets. PMID:24747743

  5. Highly multiplexed DNA sequencing by capillary electrophoresis

    SciTech Connect

    Yeung, E.S.; Ueno, K.; Chang, H.T.

    1994-12-31

    It is obvious that irrespective of whichever basic technology is eventually selected to sequence the entire human genome there are substantial gains to be made if a high degree of multiplexing of parallel runs can be implemented. Such multiplexing should not involve expensive instrumentation and should not require additional personnel, or else the main objective of cost reduction will not be satisfied even though the total time for sequencing is reduced. In the last two years, several research groups have shown that capillary electrophoresis (CE) is an attractive alternative for DNA sequencing. Part of the improvement in sequencing speed in CE is counteracted by the inherent ability of slab gels for accommodating multiple lanes in a single run. Recently, the authors have developed several excitation schemes for highly multiplexed capillary electrophoresis. Detection at the pM level was demonstrated. The authors report here the use of a novel excitation geometry to simultaneously monitor 100 capillary tubes during electrophoresis. This represents a truly parallel multiplexing scheme for high-speed DNA sequencing.

  6. Possibility of Microchip Electrophoresis for Biological Application

    NASA Astrophysics Data System (ADS)

    Kataoka, Masatoshi; Kido, Jun-Ichi; Shinohara, Yasuo

    Microchip electrophoresis has recently attracted much attention in the field of nuclear acid analysis due to its high efficiency, ease of operation, low consumption of samples and reagents, and relatively low costs. Nucleic acid fragments are separated by capillary electrophoresis in a chip with microfabricated channels, with automated detection as well as on-line data evaluation. Microfabricated devices are forecast to be fundamental to the postgenome era, especially in the field of genetics and medicine. However, although there are many reports of the use of these instruments to evaluate standard DNA, DNA ladders, PCR products, and commercially available plasmid digests, little information is available their use with biological material. In this report, we showed the accuracy of sizing and quantification of endonuclease-digested plasmid DNA. We also showed the feasibility of on-microchip endonuclease treatment of plasmid DNA and sequential analysis as an additional application for DNA analysis. Furthermore, to evaluate the possibility of microchip electrophoresis for biological application, the results of the examination of blood sugar in human plasma and mitochondrial membrane potential were shown.

  7. Darboux transformations for (1+2)-dimensional Fokker-Planck equations with constant diffusion matrix

    SciTech Connect

    Schulze-Halberg, Axel

    2012-10-15

    We construct a Darboux transformation for (1+2)-dimensional Fokker-Planck equations with constant diffusion matrix. Our transformation is based on the two-dimensional supersymmetry formalism for the Schroedinger equation. The transformed Fokker-Planck equation and its solutions are obtained in explicit form.

  8. Comparison of first dimension IPG and NEPHGE techniques in two-dimensional gel electrophoresis experiment with cytosolic unfolded protein response in Saccharomyces cerevisiae

    PubMed Central

    2013-01-01

    Background Two-dimensional gel electrophoresis (2DE) is one of the most popular methods in proteomics. Currently, most 2DE experiments are performed using immobilized pH gradient (IPG) in the first dimension; however, some laboratories still use carrier ampholytes-based isoelectric focusing technique. The aim of this study was to directly compare IPG-based and non-equilibrium pH gradient electrophoresis (NEPHGE)-based 2DE techniques by using the same samples and identical second dimension procedures. We have used commercially available Invitrogen ZOOM IPGRunner and WITAvision systems for IPG and NEPHGE, respectively. The effectiveness of IPG-based and NEPHGE-based 2DE methods was compared by analysing differential protein expression during cytosolic unfolded protein response (UPR-Cyto) in Saccharomyces cerevisiae. Results Protein loss during 2DE procedure was higher in IPG-based method, especially for basic (pI > 7) proteins. Overall reproducibility of spots was slightly better in NEPHGE-based method; however, there was a marked difference when evaluating basic and acidic protein spots. Using Coomassie staining, about half of detected basic protein spots were not reproducible by IPG-based 2DE, whereas NEPHGE-based method showed excellent reproducibility in the basic gel zone. The reproducibility of acidic proteins was similar in both methods. Absolute and relative volume variability of separate protein spots was comparable in both 2DE techniques. Regarding proteomic analysis of UPR-Cyto, the results exemplified parameters of general comparison of the methods. New highly basic protein Sis1p, overexpressed during UPR-Cyto stress, was identified by NEPHGE-based 2DE method, whereas IPG-based method showed unreliable results in the basic pI range and did not provide any new information on basic UPR-Cyto proteins. In the acidic range, the main UPR-Cyto proteins were detected and quantified by both methods. The drawback of NEPHGE-based 2DE method is its failure to

  9. Electrophoresis: The Basics (by D. M. Hawcroft)

    NASA Astrophysics Data System (ADS)

    Voige, William H.

    1999-01-01

    D. M. Hawcroft. Oxford University Press: Oxford, 1997. 142 + xii pp. Index. ISBN 0-19-963563-3. $100.00. This concise monograph is one of a series on techniques in widespread use in biochemistry and cell and molecular biology. It seeks to present, in compact and readable form, the fundamentals of electrophoresis and does so very well. Both theory and practice are included, but emphasis is on the latter. Although the preface makes it clear that this book is intended for biologists, it also deserves a place in a truly complete chemistry library. The book is logically organized. Each of the nine chapters corresponds to either a step in an electrophoresis experiment (e.g., Chapter 7: Visualization of Separated Materials) or a major application (Chapter 4: The Electrophoresis of Native and Denatured Proteins). It is written as though the reader is getting ready to begin doing electrophoresis for the first time and needs a survey of the technique and its applications. A question that occurred to me repeatedly as I read through the book is: Exactly how did the author intend it to be used? One can view the book as either a text or a laboratory manual. As a resource that might be used as a supplementary text in a graduate or upper-division undergraduate course, it does an admirable job of presenting a thorough overview of modern electrophoresis. The figures and diagrams are exceptionally clear and present useful comparisons of results that can be obtained under a variety of conditions (e.g., the resolution of DNA fragments obtained with otherwise identical wedge and normal gels). Not all its explanations, however, are as cogent. It defines how the two portions of a discontinuous gel differ but fails to explain clearly how the porosity and pH differences result in the stacking effect, which is such a gel's primary advantage. Having it on hand as a laboratory manual would be much like having colleagues who are experts in all phases of electrophoresis to consult or to go to

  10. Comparison of protein-extraction methods for gills of the shore crab, Carcinus maenas (L.), and application to 2DE.

    PubMed

    Panchout, François; Letendre, Julie; Bultelle, Florence; Denier, Xavier; Rocher, Béatrice; Chan, Philippe; Vaudry, David; Durand, Fabrice

    2013-12-01

    As it is well-established that protein extraction constitutes a crucial step for two-dimensional electrophoresis (2DE), this work was done as a prerequisite to further the study of alterations in the proteome in gills of the shore crab Carcinus maenas under contrasted environmental conditions. Because of the presence of a chitin layer, shore crab gills have an unusual structure. Consequently, they are considered as a hard tissue and represent a challenge for optimal protein extraction. In this study, we compared three published extraction procedures for subsequent applications to 2DE: the first one uses homogenization process, the second one included an additional TCA-acetone precipitation step, and finally, the third one associated grinding in liquid nitrogen (N2) and TCA-acetone precipitation. Extracted proteins were then resolved using 1DE and 2DE. Although interesting patterns were obtained using 1DE with the three methods, only the one involving grinding in liquid N2 and TCA-acetone precipitation led to proper resolution after 2DE, showing a good level of reproducibility at technical (85%) and biological (84%) levels. This last method is therefore proposed for analysis of gill proteomes in the shore crab. PMID:24294114

  11. Multiplexed Western Blotting Using Microchip Electrophoresis.

    PubMed

    Jin, Shi; Furtaw, Michael D; Chen, Huaxian; Lamb, Don T; Ferguson, Stephen A; Arvin, Natalie E; Dawod, Mohamed; Kennedy, Robert T

    2016-07-01

    Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10-20 μg per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 μm deep × 50 μm wide × 8.6 cm long channel, it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g., ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng of total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps. PMID:27270033

  12. The fluid mechanics of continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Saville, D. A.

    1990-01-01

    The overall objective is to establish theoretically and confirm experimentally the ultimate capabilities of continuous flow electrophoresis chambers operating in an environment essentially free of particle sedimentation and buoyancy. The efforts are devoted to: (1) studying the effects of particle concentration on sample conductivity and dielectric constant. The dielectric constant and conductivity were identified as playing crucial roles in the behavior of the sample and on the resolving power and throughput of continuous flow devices; and (2) improving the extant mathematical models to predict flow fields and particle trajectories in continuous flow electrophoresis. A dielectric spectrometer was designed and built to measure the complex dielectric constant of a colloidal dispersion as a function of frequency between 500 Hz and 200 kHz. The real part of the signal can be related to the sample's conductivity and the imaginary part to its dielectric constant. Measurements of the dielectric constants of several different dispersions disclosed that the dielectric constants of dilute systems of the sort encountered in particle electrophoresis are much larger than would be expected based on the extant theory. Experiments were carried out to show that, in many cases, this behavior is due to the presence of a filamentary structure of small hairs on the particle surface. A technique for producing electrokinetically ideal synthetic latex particles by heat treating was developed. Given the ubiquitous nature of hairy surfaces with both cells and synthetic particles, it was deemed necessary to develop a theory to explain their behavior. A theory for electrophoretic mobility of hairy particles was developed. Finally, the extant computer programs for predicting the structure of electro-osmotically driven flows were extended to encompass flow channels with variable wall mobilities.

  13. Serum protein electrophoresis in spontaneous canine hyperadrenocorticalism.

    PubMed

    van den Broek, A H; Lida, J

    1989-01-01

    The serum protein concentrations of dogs with confirmed spontaneous hyperadrenocorticalism were determined by agarose gel electrophoresis before and during treatment with mitotane. In untreated animals a significant increase was detected in the mean concentration of total protein and the mean concentration and percentage of alpha-2 globulin. The mean concentration and percentage of albumin and gamma-globulin were significantly decreased. In animals on treatment the mean concentration of total proteins and the mean concentration and percentage of beta-2 globulin were significantly reduced. PMID:2466309

  14. Pulsed field gel electrophoresis for dairy propionibacteria.

    PubMed

    Chuat, Victoria; de Freitas, Rosangela; Dalmasso, Marion

    2015-01-01

    Pulsed field gel electrophoresis (PFGE) is a technique using alternating electric fields to migrate high molecular weight DNA fragments with a high resolution. This method consists of the digestion of bacterial chromosomal DNA with rare-cutting restriction enzymes and in applying an alternating electrical current between spatially distinct pairs of electrodes. DNA molecules migrate at different speeds according to the size of the fragments. Among other things, this technique is considered as the "gold standard" for genotyping, genetic fingerprinting, epidemiological studies, genome size estimation, and studying radiation-induced DNA damage and repair. This chapter describes a PFGE method that can be used to differentiate dairy propionibacteria. PMID:25862063

  15. Microfabricated capillary array electrophoresis device and method

    DOEpatents

    Simpson, Peter C.; Mathies, Richard A.; Woolley, Adam T.

    2004-06-15

    A capillary array electrophoresis (CAE) micro-plate with an array of separation channels connected to an array of sample reservoirs on the plate. The sample reservoirs are organized into one or more sample injectors. One or more waste reservoirs are provided to collect wastes from reservoirs in each of the sample injectors. Additionally, a cathode reservoir is also multiplexed with one or more separation channels. To complete the electrical path, an anode reservoir which is common to some or all separation channels is also provided on the micro-plate. Moreover, the channel layout keeps the distance from the anode to each of the cathodes approximately constant.

  16. Microfabricated capillary array electrophoresis device and method

    DOEpatents

    Simpson, Peter C.; Mathies, Richard A.; Woolley, Adam T.

    2000-01-01

    A capillary array electrophoresis (CAE) micro-plate with an array of separation channels connected to an array of sample reservoirs on the plate. The sample reservoirs are organized into one or more sample injectors. One or more waste reservoirs are provided to collect wastes from reservoirs in each of the sample injectors. Additionally, a cathode reservoir is also multiplexed with one or more separation channels. To complete the electrical path, an anode reservoir which is common to some or all separation channels is also provided on the micro-plate. Moreover, the channel layout keeps the distance from the anode to each of the cathodes approximately constant.

  17. Cytokine Analysis by Immunoaffinity Capillary Electrophoresis

    PubMed Central

    Mendonca, Mark; Kalish, Heather

    2014-01-01

    Immunoaffinity capillary electrophoresis (ICE) is a powerful tool used to detect and quantify target proteins of interest in complex biological fluids. The target analyte is captured and bound to antibodies immobilized onto the wall of a capillary, labeled in situ with a fluorescent dye, eluted and detected online using laser-induced fluorescence following electrophoretic separation. Here, we illustrate how to construct an immunoaffinity capillary and utilize it to run ICE in order to capture and quantify target cytokines and chemokines from a clinical sample. PMID:22976107

  18. Hybrid slab-microchannel gel electrophoresis system

    DOEpatents

    Balch, Joseph W.; Carrano, Anthony V.; Davidson, James C.; Koo, Jackson C.

    1998-01-01

    A hybrid slab-microchannel gel electrophoresis system. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate.

  19. Analysis of Stevia glycosides by capillary electrophoresis.

    PubMed

    Mauri, P; Catalano, G; Gardana, C; Pietta, P

    1996-02-01

    The determination of diterpene glycosides from Stevia rebaudiana leaves using capillary electrophoresis is described. Analyses were performed on fused silica capillaries with 20 mM sodium tetraborate buffer, pH 8.3, and 30 mM sodium dodecyl sulfate. The effect of the organic solvent injected with the sample solution on the electrophoretic solution has been confirmed, and an absolute amount of 1.6 nL per injected sample was optimal. Rebaudioside A and steviolbioside were isolated by semipreparative high performance liquid chromatography (HPLC), and their structure was assessed by mass spectrometry. PMID:8900944

  20. Gravitational Fields with 2-Dimensional Killing Leaves and the Gravitational Interaction of Light

    NASA Astrophysics Data System (ADS)

    Vilasi, Gaetano

    Gravitational fields invariant for a non Abelian Lie algebra generating a 2-dimensional distribution, are explicitly described. When the orthogonal distribution is integrable and the metric is not degenerate along the orbits, these solutions are parameterized either by solutions of a transcendental equation (the tortoise equation), or by solutions of Darboux equation. Metrics, corresponding to solutions of the tortoise equation, are characterized as those that admit a 3-dimensional Lie algebra of Killing fields with 2-dimensional leaves. It is shown that the remaining metrics represent nonlinear gravitational waves obeying to two nonlinearsuperposition laws. The energy and the polarization of this family of waves are explicitly evaluated; it is shown that they have spin-1 and their possible sources are also described. Old results by Tolman, Ehrenfest, Podolsky and Wheeler on the gravitational interaction of photons are naturally reinterpreted.

  1. Analysis of Protein Oligomerization by Electrophoresis.

    PubMed

    Cubillos-Rojas, Monica; Schneider, Taiane; Sánchez-Tena, Susana; Bartrons, Ramon; Ventura, Francesc; Rosa, Jose Luis

    2016-01-01

    A polypeptide chain can interact with other polypeptide chains and form stable and functional complexes called "oligomers." Frequently, biochemical analysis of these complexes is made difficult by their great size. Traditionally, size exclusion chromatography, immunoaffinity chromatography, or immunoprecipitation techniques have been used to isolate oligomers. Components of these oligomers are then further separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and identified by immunoblotting with specific antibodies. Although they are sensitive, these techniques are not easy to perform and reproduce. The use of Tris-acetate polyacrylamide gradient gel electrophoresis allows the simultaneous analysis of proteins in the mass range of 10-500 kDa. We have used this characteristic together with cross-linking reagents to analyze the oligomerization of endogenous proteins with a single electrophoretic gel. We demonstrate how the oligomerization of p53, the pyruvate kinase isoform M2, or the heat shock protein 27 can be studied with this system. We also show how this system is useful for studying the oligomerization of large proteins such as clathrin heavy chain or the tuberous sclerosis complex. Oligomerization analysis is dependent on the cross-linker used and its concentration. All of these features make this system a very helpful tool for the analysis of protein oligomerization. PMID:27613048

  2. Capillary electrophoresis of DNA for molecular diagnostics.

    PubMed

    Righetti, P G; Gelfi, C

    1997-09-01

    A number of applications of capillary zone electrophoresis (CZE) in sieving liquid polymers (notably linear polyacrylamides and cellulose) for the analysis of polymerase chain reaction (PCR) products of clinically relevant, diagnostic DNA, are reviewed. The fields covered are: human genetics, quantitative gene dosage, microbiology and virology, forensic medicine and therapeutic DNA (notably, antisense nucleotides). Some unique, novel developments are highlighted, such as: (i) nonisocratic CZE, i.e., temperature-programmed CZE for detection of DNA point mutations; (ii) the synthesis of novel N-substituted acrylamides, offering extreme resistance to alkaline hydrolysis coupled to high hydrophilicity. In the field of denaturing gradient gel electrophoresis (DGGE), as routinely performed in gel slabs, a novel methodology is described in CZE: double-gradient DGGE. In this technique, two gradients are simultaneously applied along the migration direction: a chemical (or thermal) denaturing gradient, for partially unwinding homo- and hetero-duplexes of DNA, and a porosity gradient, for recompacting diffuse bands melting over a broader range of denaturing conditions. It is thus demonstrated that chemical gradients, in addition to temperature gradients, can be easily implemented even in a capillary format. PMID:9372261

  3. Gel Electrophoresis of Gold-DNA Nanoconjugates

    DOE PAGESBeta

    Pellegrino, T.; Sperling, R. A.; Alivisatos, A. P.; Parak, W. J.

    2007-01-01

    Gold-DNA conjugates were investigated in detail by a comprehensive gel electrophoresis study based on 1200 gels. A controlled number of single-stranded DNA of different length was attached specifically via thiol-Au bonds to phosphine-stabilized colloidal gold nanoparticles. Alternatively, the surface of the gold particles was saturated with single stranded DNA of different length either specifically via thiol-Au bonds or by nonspecific adsorption. From the experimentally determined electrophoretic mobilities, estimates for the effective diameters of the gold-DNA conjugates were derived by applying two different data treatment approaches. The first method is based on making a calibration curve for the relation between effectivemore » diameters and mobilities with gold nanoparticles of known diameter. The second method is based on Ferguson analysis which uses gold nanoparticles of known diameter as reference database. Our study shows that effective diameters derived from gel electrophoresis measurements are affected with a high error bar as the determined values strongly depend on the method of evaluation, though relative changes in size upon binding of molecules can be detected with high precision. Furthermore, in this study, the specific attachment of DNA via gold-thiol bonds to Au nanoparticles is compared to nonspecific adsorption of DNA. Also, the maximum number of DNA molecules that can be bound per particle was determined.« less

  4. Capillary zone electrophoresis of large DNA.

    PubMed

    Guszczynski, T; Pulyaeva, H; Tietz, D; Garner, M M; Chrambach, A

    1993-01-01

    Capillary zone electrophoresis (CZE) of DNA 23.1 to 48.5 kb in length in polyacrylamide solutions of several concentrations provides evidence for polymer concentration and DNA length-dependent stretching and orientation of these species and suggests an effective separation at a polymer concentration of about 0.6%. Applying a 0.1% polyacrylamide concentration to the lambda-phage DNA ladder, at least 5 components are separated; separation improves with lowering of the field strength to 2 V/cm and, correspondingly, extended duration of CZE. Saccharomyces pombe chromosomal DNA separates into 3 major components on CZE at high field strength (270 V/cm) in 0.9% polyacrylamide solution, confirming a previous finding made on electrophoresis in a 1.1 mm ID tube at low field strength. However, the finding is limited to one source of the DNA plug, and the chromosomal identity of the components remains unknown. Methodological problems in the CZE of large DNA relate to the need for extended duration of pressure injection if absorbance detection is applied, the need to define the starting zone after extended pressure injection, the need to melt and digest agarose plugs prior to loading, and related needs for thermostating of the sample chamber and for software compatible with low voltage operation. PMID:8354238

  5. Determination of surfactants by capillary electrophoresis.

    PubMed

    Heinig, K; Vogt, C

    1999-10-01

    Capillary electrophoresis has been increasingly used during the past few years for the separation and determination of surfactants. These substances are applied in many household and industrial products such as laundry detergents, cosmetics and pharmaceuticals, often as homologous and isomeric mixtures. Product development and control as well as toxicological and environmental analyses require selective and sensitive analytical methods. This review presents capillary electrophoretic techniques to determine important representatives of cationic, anionic, and neutral surfactants. The application of different buffer additives such as organic solvents, cyclodextrins or micelles to enhance the resolution of complex mixtures is discussed. Besides direct and indirect UV and fluorescence detection, examples for conductivity and mass spectrometric detection are also given. Derivatization procedures to improve the detectability and implement charge in neutral analytes are described. The successful use of capillary electrophoresis for surfactant determinations has proven that it can serve as a routine technique in many real-world applications. Robust, validated methods for the quantitation of single compounds, such as alkylbenzene sulfonates, sodium dodecyl sulfate and benzalkonium salts, are now available. Characteristic peak patterns (fingerprint analysis) can be used for the identification of surfactants in multicomponent formulations (e.g. ethoxylates and phosphonates). PMID:10596832

  6. Capillary electrophoresis coupled with automated fraction collection.

    PubMed

    Huge, Bonnie Jaskowski; Flaherty, Ryan J; Dada, Oluwatosin O; Dovichi, Norman J

    2014-12-01

    A fraction collector based on a drop-on-demand ink-jet printer was developed to interface capillary zone electrophoresis with a 96 well microtiter plate. We first evaluated the performance of the collector by using capillary zone electrophoresis to analyze a 1mM solution of tetramethylrhodamine; a fluorescent microtiter plate reader was then used to detect the analyte and characterize fraction carryover between wells. Relative standard deviation in peak height was 20% and the relative standard deviation in migration time was 1%. The mean and standard deviation of the tetramethylrhodamine peak width was 5 ± 1 s and likely limited by the 4-s period between droplet deposition. We next injected a complex mixture of DNA fragments and used real-time PCR to quantify the product in a CE-SELEX experiment. The reconstructed electrophoretic peak was 27 s in duration. Finally, we repeated the experiment in the presence of a 30-µM thrombin solution under CE-SELEX conditions; fractions were collected and next-generation sequencing was used to characterize the DNA binders. Over 25,000 sequences were identified with close matches to known thrombin binding aptamers. PMID:25159411

  7. Using capillary electrophoresis to characterize polymeric particles.

    PubMed

    Riley, Kathryn R; Liu, Sophia; Yu, Guo; Libby, Kara; Cubicciotti, Roger; Colyer, Christa L

    2016-09-01

    Capillary electrophoresis (CE) was used for the characterization of a variety of polymeric micron and sub-micron particles based on size, surface functionality, and binding properties. First, a robust capillary zone electrophoresis (CZE) method was developed for the baseline separation and quantitation of commercially available polystyrene particles with various surface modifications (including amino, carboxylate, and sulfate functional groups) and various sizes (0.2, 0.5, 1.0, and 3.0μm). The separation of DNA-templated polyacrylamide particles from untemplated particles (as used for the Ion Torrent Personal Genome Machine) was demonstrated. Finally, using the 29-base thrombin aptamer and thrombin protein as a model system, a study was undertaken to determine dissociation constants for the aptamer and protein in free solution and when the aptamer was conjugated to a particle, with the goal of better understanding how the use of solid substrates, like particles, affects selection and binding processes. Dissociation constants were determined and were found to be approximately 5-fold higher for the aptamer conjugated to a particle relative to that in free solution. PMID:27543386

  8. Capillary Electrophoresis coupled with Automated Fraction Collection

    PubMed Central

    Huge, Bonnie Jaskowski; Flaherty, Ryan; Dada, Oluwatosin O.; Dovichi, Norman J.

    2014-01-01

    A fraction collector based on a drop-on-demand ink-jet printer was developed to interface capillary zone electrophoresis with a 96 well microtiter plate. We first evaluated the performance of the collector by using capillary zone electrophoresis to analyze a 1 mM solution of tetramethylrhodamine; a fluorescent microtiter plate reader was then used to detect the analyte and characterize fraction carryover between wells. Relative standard deviation in peak height was 20% and the relative standard deviation in migration time was 1%. The mean and standard deviation of the tetramethylrhodamine peak width was 5 ± 1 s and likely limited by the 4-s period between droplet deposition. We next injected a complex mixture of DNA fragments and used real-time PCR to quantify the product in a CE-SELEX experiment. The reconstructed electrophoretic peak was 27 s in duration. Finally, we repeated the experiment in the presence of a 30-μM thrombin solution under CE-SELEX conditions; fractions were collected and next-generation sequencing was used to characterize the DNA binders. Over 25,000 sequences were identified with close matches to known thrombin binding aptamers. PMID:25159411

  9. Fractionation of mineral species by electrophoresis

    NASA Technical Reports Server (NTRS)

    Dunning, J. D.; Herren, B. J.; Tipps, R. W.; Snyder, R. S.

    1982-01-01

    The fractionation of fine-grained aggregates into their major components is a problem in many scientific areas including earth and planetary science. Electrophoresis, the transport of electrically charged particles, immersed in a suspension medium, by a direct current field (Bier, 1959), was employed in this study as a means of separating simulated lunar soil into its constituent minerals. In these tests, conducted in a static analytical cylindrical microelectrophoresis apparatus, samples of simulated lunar soil and samples of pure mineral constituents were placed in the chamber; the electrophoretic mobilities of the lunar soil and the individual mineral constituents were measured. In most of the suspension buffers employed separability was indicated, on the basis of differences in mobility, for all the constituent mineral species except ilmenite and pyroxene, which were not efficiently separable in any of the buffers. Although only a few suspension media were employed, the success of this initial study suggests that electrophoresis may be an important mineral fractionation option in fine-grained aggregate processing.

  10. A microchannel electrophoresis DNA sequencing system

    SciTech Connect

    Madabhushi, R S; Warth, T; Balch, J W; Bass, M; Brewer, L R; Copeland, A C; Davidson, J C; Fitch, J P; Kegelmeyer, L M; Kimbrough, J R; McCready, P; Nelson, D; Pastrone, R L; Richardson, P M; Swierkowski, S P; Tarte, L A; Vainer, M

    1999-01-01

    In order to increase the DNA sequencing throughput of the Joint Genome Institute, we have developed a microchannel electrophoresis system. The critical new and unique elements of this system include 1) a process for the production of arrays of 96 and 384 microchannels on bonded glass substrates up to 14 x 58 cm and 2) new sieving media for high resolution and high speed separations. With custom fabrication apparatus, microchannels are etched in a borosilicate substrate, and then fusion bonded to a top substrate 1.1 mm thick that has access holes formed in it. SEM examination shows a typical microchannel to be 40 micrometers deep x 180 micrometers wide by 46 cm long. This technology offers significant advantages over discrete capillaries or conventional slab-gel approaches. High throughput DNA sequencing with over 550 base pairs resolution has been achieved in roughly half the time of conventional sequencers. In February 1999, we begin a pre-production evaluation protocol for the microchannel and for three glass capillary electrophoresis systems (two from industry and one developed by Lawrence Berkeley National Laboratory for the Joint Genome Institute). In order to utilize these instruments for DNA production sequencing, we have been evaluating and implementing software to convert raw electropherograms into called DNA bases with an associated probability of error. Our original intent was to utilize the DNA base calling software known as Plan and Phred developed by the University of Washington. This software has been outstanding for our slab gel electrophoresis systems currently in the production facility. In our tests and evaluations of this software applied to microchannel data, we observed that the electropherograms are of a different statistical and underlying signal structure compared to slab gels. Even with substantial modifications to the software, base calling performance was not satisfactory for the microchannel data. In this paper, we will present o The

  11. 2-DE using hemi-fluorinated surfactants.

    PubMed

    Starita-Geribaldi, Mireille; Thebault, Pascal; Taffin de Givenchy, Elisabeth; Guittard, Frederic; Geribaldi, Serge

    2007-07-01

    The synthesis of hemi-fluorinated zwitterionic surfactants was realized and assessed for 2-DE, a powerful separation method for proteomic analysis. These new fluorinated amidosulfobetaine (FASB-p,m) were compared to their hydrocarbon counterparts amidosulfobetaine (ASB-n) characterized by a hydrophilic polar head, a hydrophobic and lipophilic tail, and an amido group as connector. The tail of these FASB surfactants was in part fluorinated resulting in the modulation of its lipophilicity (or oleophobicity). Their effect on the red blood cell (RBC) membrane showed a specific solubilization depending on the length of the hydrophobic part. A large number of polypeptide spots appeared in the 2-DE patterns by using FASB-p,m. The oleophobic character of these surfactants was confirmed by the fact that Band 3, a highly hydrophobic transmembrane protein, was not solubilized by these fluorinated structures. The corresponding pellet was very rich in Band 3 and could then be solubilized by using a strong detergent such as amidosulfobetaine with an alkyl tail containing 14 carbon atoms (ASB-14). Thus, these hemi-fluorinated surfactants appeared as powerful tools when used at the first step of a two-step solubilization strategy using a hydrocarbon homologous surfactant in the second step. PMID:17577887

  12. Phosphoproteome profiling using a fluorescent phosphosensor dye in two-dimensional polyacrylamide gel electrophoresis.

    PubMed

    Otani, Mieko; Taniguchi, Taizo; Sakai, Akiko; Seta, Jouji; Kadoyama, Keiichi; Nakamura-Hirota, Tooru; Matsuyama, Shogo; Sano, Keiji; Takano, Masaoki

    2011-07-01

    We validated the novel PhosphoQUANTI SolidBlue Complex (PQSC) dye for the sensitive fluorescent detection of phosphorylated proteins in polyacrylamide- and two-dimensional gel electrophoresis (PAGE and 2DE, respectively). PQSC can detect as little as 15.6 ng of ß-casein, a pentaphosphorylated protein, and 61.3 ng of ovalbumin, a diphosphorylated protein. Fluorescence intensity correlates with the number of phosphorylated residues on the protein. To demonstrate the specificity of PQSC for phosphoproteins, enzymatically dephosphorylated lysates of Swiss 3T3 cells were separated in 2DE gels and stained by PQSC. The fluorescence signals in these gels were markedly reduced following dephosphorylation. When the phosphorylated proteins in Swiss 3T3 cell lysates were concentrated using a phosphoprotein enrichment column, the majority of phosphoproteins showed fluorescence signals in the pI 4-5 range. Finally, we performed phosphoproteome analysis to study differences in the protein phosphorylation profiles of proliferating and quiescent Swiss 3T3 cells. Over 135 discernible protein spots were detected, from which a selection of 15 spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF-MS). The PQSC staining procedure for phosphoprotein detection is simple, reversible, and fully compatible with MALDI TOF-MS. PMID:21384102

  13. Automated DNA electrophoresis, hybridization and detection

    SciTech Connect

    Zapolski, E.J.; Gersten, D.M.; Golab, T.J.; Ledley, R.S.

    1986-05-01

    A fully automated, computer controlled system for nucleic acid hybridization analysis has been devised and constructed. In practice, DNA is digested with restriction endonuclease enzyme(s) and loaded into the system by pipette; /sup 32/P-labelled nucleic acid probe(s) is loaded into the nine hybridization chambers. Instructions for all the steps in the automated process are specified by answering questions that appear on the computer screen at the start of the experiment. Subsequent steps are performed automatically. The system performs horizontal electrophoresis in agarose gel, fixed the fragments to a solid phase matrix, denatures, neutralizes, prehybridizes, hybridizes, washes, dries and detects the radioactivity according to the specifications given by the operator. The results, printed out at the end, give the positions on the matrix to which radioactivity remains hybridized following stringent washing.

  14. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, R.D.; Udseth, H.R.; Olivares, J.A.

    1994-10-18

    A system and method for analyzing molecular constituents of a composition sample include: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5--100 kVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g.,{+-}2--8 kVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit. 21 figs.

  15. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, R.P.; Udseth, H.R.; Olivares, J.A.

    1989-12-05

    A system and method for analyzing molecular constituents of a composition sample includes: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5--100 kVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g., [+-]2--8 kVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit. 21 figs.

  16. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, Richard P.; Udseth, Harold R.; Olivares, Jose A.

    1989-01-01

    A system and method for analyzing molecular constituents of a composition sample includes: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5-100 KVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g., .+-.2-8 KVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit.

  17. Combined electrophoresis-electrospray interface and method

    DOEpatents

    Smith, Richard D.; Udseth, Harold R.; Olivares, Jose A.

    1994-10-18

    A system and method for analyzing molecular constituents of a composition sample includes: forming a solution of the sample, separating the solution by capillary electrophoresis into an eluent of constituents longitudinally separated according to their relative electrophoretic mobilities, electrospraying the eluent to form a charged spray in which the molecular constituents have a temporal distribution; and detecting or collecting the separated constituents in accordance with the temporal distribution in the spray. A first high-voltage (e.g., 5-100 KVDC) is applied to the solution. The spray is charged by applying a second high voltage (e.g., .+-.2-8 KVDC) between the eluent at the capillary exit and a cathode spaced in front of the exit. A complete electrical circuit is formed by a conductor which directly contacts the eluent at the capillary exit, or by conduction through a sheath electrode discharged in an annular sheath flow about the capillary exit.

  18. Hybrid slab-microchannel gel electrophoresis system

    DOEpatents

    Balch, J.W.; Carrano, A.V.; Davidson, J.C.; Koo, J.C.

    1998-05-05

    A hybrid slab-microchannel gel electrophoresis system is described. The hybrid system permits the fabrication of isolated microchannels for biomolecule separations without imposing the constraint of a totally sealed system. The hybrid system is reusable and ultimately much simpler and less costly to manufacture than a closed channel plate system. The hybrid system incorporates a microslab portion of the separation medium above the microchannels, thus at least substantially reducing the possibility of non-uniform field distribution and breakdown due to uncontrollable leakage. A microslab of the sieving matrix is built into the system by using plastic spacer materials and is used to uniformly couple the top plate with the bottom microchannel plate. 4 figs.

  19. Applications of capillary electrophoresis in biotechnology.

    PubMed

    Lagu, A L

    1999-10-01

    Capillary electrophoresis (CE)-related techniques are increasingly being used as a matter of routine practice in the biotechnology discipline. Since recombinant DNA-derived proteins and the antisense oligonucleotides constitute a large portion of the applications of these techniques, they have been emphasized in this review. Analyses by CE of Escherichia coli-derived proteins and glycosylated proteins derived from mammalian cell cultures are summarized, as well as those of the carbohydrate chains that have been enzymatically removed from the protein. Applications of CE in the analysis of the antisense oligonucleotides for the determination of purity and the analytical studies on the metabolism of these modified oligonucleotides, by CE are reviewed. The literature mainly covers the period from 1996. PMID:10596822

  20. Novel absorption detection techniques for capillary electrophoresis

    SciTech Connect

    Xue, Y.

    1994-07-27

    Capillary electrophoresis (CE) has emerged as one of the most versatile separation methods. However, efficient separation is not sufficient unless coupled to adequate detection. The narrow inner diameter (I.D.) of the capillary column raises a big challenge to detection methods. For UV-vis absorption detection, the concentration sensitivity is only at the {mu}M level. Most commercial CE instruments are equipped with incoherent UV-vis lamps. Low-brightness, instability and inefficient coupling of the light source with the capillary limit the further improvement of UV-vis absorption detection in CE. The goals of this research have been to show the utility of laser-based absorption detection. The approaches involve: on-column double-beam laser absorption detection and its application to the detection of small ions and proteins, and absorption detection with the bubble-shaped flow cell.

  1. Antibody enhancement of free-flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Cohly, H. H. P.; Morrison, Dennis R.; Atassi, M. Zouhair

    1988-01-01

    Specific T cell clones and antibodies (ABs) were developed to study the efficiency of purifying closely associated T cells using Continuous Flow Electrophoresis System. Enhanced separation is accomplished by tagging cells first with ABs directed against the antigenic determinants on the cell surface and then with ABs against the Fc portion of the first AB. This second AB protrudes sufficiently beyond the cell membrane and glycocalyx to become the major overall cell surface potential determinant and thus causes a reduction of electrophoretic mobility. This project was divided into three phases. Phase one included development of specific T cell clones and separation of these specific clones. Phase two extends these principles to the separation of T cells from spleen cells and immunized lymph node cells. Phase three applies this double antibody technique to the separation of T cytotoxic cells from bone marrow.

  2. Analysis of starch structure using fluorophore-assisted carbohydrate electrophoresis.

    PubMed

    Morell, M K; Samuel, M S; O'Shea, M G

    1998-11-01

    The analysis of the fine structure of starches is important to the investigation of linkages between starch structure and function and to the investigation of the properties and roles of starch biosynthetic, modifying and degradation enzymes. Fluorophore-assisted carbohydrate electrophoresis has recently been introduced as a method for the analysis of the oligosaccharide populations released by the enzymatic digestion of starches, which has advantages in resolution and sensitivity over previously used methods, and provides the capacity for the facile analysis of oligosaccharide populations on either a molar or mass basis. The use of fluorophore-assisted carbohydrate electrophoresis for the analysis of oligosaccharides is reviewed with particular reference to the choice of label, efficiency of labeling and separation techniques. Examples of separations using slab gel electrophoresis, DNA sequencer analysis and capillary electrophoresis are presented and we conclude that on the basis of resolution and reproducibility, capillary electrophoresis is the method of choice for the separation of oligosaccharides of degree of polymerization from 1 to 100. Examples of isoamylase-debranched starches and glycogens analyzed by capillary electrophoresis are presented. The capillary electrophoresis analysis of starch structure through the analysis of oligosaccharides released by the debranching of limit dextrins derived from starches and glycogens is introduced as a useful diagnostic of starch structure. The potential for future development of novel diagnostics for starch structure using fluorophore-assisted carbohydrate electrophoresis is discussed. PMID:9848667

  3. Inexpensive and Safe DNA Gel Electrophoresis Using Household Materials

    ERIC Educational Resources Information Center

    Ens, S.; Olson, A. B.; Dudley, C.; Ross, N. D., III; Siddiqi, A. A.; Umoh, K. M.; Schneegurt, M. A.

    2012-01-01

    Gel electrophoresis is the single most important molecular biology technique and it is central to life sciences research, but it is often too expensive for the secondary science classroom or homeschoolers. A simple safe low-cost procedure is described here that uses household materials to construct and run DNA gel electrophoresis. Plastic…

  4. Gel Electrophoresis on a Budget to Dye for

    ERIC Educational Resources Information Center

    Yu, Julie H.

    2010-01-01

    Gel electrophoresis is one of the most important tools used in molecular biology and has facilitated the entire field of genetic engineering by enabling the separation of nucleic acids and proteins. However, commercial electrophoresis kits can cost up to $800 for each setup, which is cost prohibitive for most classroom budgets. This article…

  5. Damage spreading in 2-dimensional isotropic and anisotropic Bak-Sneppen models

    NASA Astrophysics Data System (ADS)

    Bakar, B.; Tirnakli, U.

    2008-03-01

    We implement the damage spreading technique on 2-dimensional isotropic and anisotropic Bak-Sneppen models. Our extensive numerical simulations show that there exists a power-law sensitivity to the initial conditions at the statistically stationary state (self-organized critical state). Corresponding growth exponent α for the Hamming distance and the dynamical exponent z are calculated. These values allow us to observe a clear data collapse of the finite size scaling for both versions of the Bak-Sneppen model. Moreover, it is shown that the growth exponent of the distance in the isotropic and anisotropic Bak-Sneppen models is strongly affected by the choice of the transient time.

  6. Dynamical analysis and simulation of a 2-dimensional disease model with convex incidence

    NASA Astrophysics Data System (ADS)

    Yu, Pei; Zhang, Wenjing; Wahl, Lindi M.

    2016-08-01

    In this paper, a previously developed 2-dimensional disease model is studied, which can be used for both epidemiologic modeling and in-host disease modeling. The main attention of this paper is focused on various dynamical behaviors of the system, including Hopf and generalized Hopf bifurcations which yield bistability and tristability, Bogdanov-Takens bifurcation, and homoclinic bifurcation. It is shown that the Bogdanov-Takens bifurcation and homoclinic bifurcation provide a new mechanism for generating disease recurrence, that is, cycles of remission and relapse such as the viral blips observed in HIV infection.

  7. A Finger-Shaped Tactile Sensor for Fabric Surfaces Evaluation by 2-Dimensional Active Sliding Touch

    PubMed Central

    Hu, Haihua; Han, Yezhen; Song, Aiguo; Chen, Shanguang; Wang, Chunhui; Wang, Zheng

    2014-01-01

    Sliding tactile perception is a basic function for human beings to determine the mechanical properties of object surfaces and recognize materials. Imitating this process, this paper proposes a novel finger-shaped tactile sensor based on a thin piezoelectric polyvinylidene fluoride (PVDF) film for surface texture measurement. A parallelogram mechanism is designed to ensure that the sensor applies a constant contact force perpendicular to the object surface, and a 2-dimensional movable mechanical structure is utilized to generate the relative motion at a certain speed between the sensor and the object surface. By controlling the 2-dimensional motion of the finger-shaped sensor along the object surface, small height/depth variation of surface texture changes the output charge of PVDF film then surface texture can be measured. In this paper, the finger-shaped tactile sensor is used to evaluate and classify five different kinds of linen. Fast Fourier Transformation (FFT) is utilized to get original attribute data of surface in the frequency domain, and principal component analysis (PCA) is used to compress the attribute data and extract feature information. Finally, low dimensional features are classified by Support Vector Machine (SVM). The experimental results show that this finger-shaped tactile sensor is effective and high accurate for discriminating the five textures. PMID:24618775

  8. Determining the Best Sensing Coverage for 2-Dimensional Acoustic Target Tracking

    PubMed Central

    Pashazadeh, Saeid; Sharifi, Mohsen

    2009-01-01

    Distributed acoustic target tracking is an important application area of wireless sensor networks. In this paper we use algebraic geometry to formally model 2-dimensional acoustic target tracking and then prove its best degree of required sensing coverage. We present the necessary conditions for three sensing coverage to accurately compute the spatio-temporal information of a target object. Simulations show that 3-coverage accurately locates a target object only in 53% of cases. Using 4-coverage, we present two different methods that yield correct answers in almost all cases and have time and memory usage complexity of Θ(1). Analytic 4-coverage tracking is our first proposed method that solves a simultaneous equation system using the sensing information of four sensor nodes. Redundant answer fusion is our second proposed method that solves at least two sets of simultaneous equations of target tracking using the sensing information of two different sets of three sensor nodes, and fusing the results using a new customized formal majority voter. We prove that 4-coverage guarantees accurate 2-dimensional acoustic target tracking under ideal conditions. PMID:22412319

  9. A finger-shaped tactile sensor for fabric surfaces evaluation by 2-dimensional active sliding touch.

    PubMed

    Hu, Haihua; Han, Yezhen; Song, Aiguo; Chen, Shanguang; Wang, Chunhui; Wang, Zheng

    2014-01-01

    Sliding tactile perception is a basic function for human beings to determine the mechanical properties of object surfaces and recognize materials. Imitating this process, this paper proposes a novel finger-shaped tactile sensor based on a thin piezoelectric polyvinylidene fluoride (PVDF) film for surface texture measurement. A parallelogram mechanism is designed to ensure that the sensor applies a constant contact force perpendicular to the object surface, and a 2-dimensional movable mechanical structure is utilized to generate the relative motion at a certain speed between the sensor and the object surface. By controlling the 2-dimensional motion of the finger-shaped sensor along the object surface, small height/depth variation of surface texture changes the output charge of PVDF film then surface texture can be measured. In this paper, the finger-shaped tactile sensor is used to evaluate and classify five different kinds of linen. Fast Fourier Transformation (FFT) is utilized to get original attribute data of surface in the frequency domain, and principal component analysis (PCA) is used to compress the attribute data and extract feature information. Finally, low dimensional features are classified by Support Vector Machine (SVM). The experimental results show that this finger-shaped tactile sensor is effective and high accurate for discriminating the five textures. PMID:24618775

  10. Validation of STR typing by capillary electrophoresis.

    PubMed

    Moretti, T R; Baumstark, A L; Defenbaugh, D A; Keys, K M; Brown, A L; Budowle, B

    2001-05-01

    With the use of capillary electrophoresis (CE), high-resolution electrophoretic separation of short tandem repeat (STR) loci can be achieved in a semiautomated fashion. Laser-induced detection of fluorescently labeled PCR products and multicolor analysis enable the rapid generation of multilocus DNA profiles. In this study, conditions for typing PCR-amplified STR loci by capillary electrophoresis were investigated using the ABI Prism 310 Genetic Analyzer (Applied Biosystems). An internal size standard was used with each run to effectively normalize mobility differences among injections. Alleles were designated by comparison to allelic ladders that were run with each sample set. Multiple runs of allelic ladders and of amplified samples demonstrate that allele sizes were reproducible, with standard deviations typically less than 0.12 bases for fragments up to 317 bases in length (largest allele analyzed) separated in a 47 cm capillary. Therefore, 99.7% of all alleles that are the same length should fall within the measurement error window of +/- 0.36 bases. Microvariants of the tetranucleotide repeats were also accurately typed by the analytical software. Alleles differing in size by one base could be resolved in two-donor DNA mixtures in which the minor component comprised > or = 5% of the total DNA. Furthermore, the quantitative data format (i.e., peak amplitude) can in some instances assist in determining individual STR profiles in mixed samples. DNA samples from previously typed cases (typed for RFLP, AmpliType PM+DQA1, and/or D1S80) were amplified using AmpFlSTR Profiler Plus and COfiler and were evaluated using the ABI Prism 310. Most samples yielded typable results. Compared with previously determined results for other loci, there were no discrepancies as to the inclusion or exclusion of suspects or victims. CE thus provides efficient separation, resolution, sensitivity and precision, and the analytical software provides reliable genotyping of STR loci. The

  11. The PROTICdb database for 2-DE proteomics.

    PubMed

    Langella, Olivier; Zivy, Michel; Joets, Johann

    2007-01-01

    PROTICdb is a web-based database mainly designed to store and analyze plant proteome data obtained by 2D polyacrylamide gel electrophoresis (2D PAGE) and mass spectrometry (MS). The goals of PROTICdb are (1) to store, track, and query information related to proteomic experiments, i.e., from tissue sampling to protein identification and quantitative measurements; and (2) to integrate information from the user's own expertise and other sources into a knowledge base, used to support data interpretation (e.g., for the determination of allelic variants or products of posttranslational modifications). Data insertion into the relational database of PROTICdb is achieved either by uploading outputs from Mélanie, PDQuest, IM2d, ImageMaster(tm) 2D Platinum v5.0, Progenesis, Sequest, MS-Fit, and Mascot software, or by filling in web forms (experimental design and methods). 2D PAGE-annotated maps can be displayed, queried, and compared through the GelBrowser. Quantitative data can be easily exported in a tabulated format for statistical analyses with any third-party software. PROTICdb is based on the Oracle or the PostgreSQLDataBase Management System (DBMS) and is freely available upon request at http://cms.moulon.inra.fr/content/view/14/44/. PMID:17093318

  12. Electrolytic Reduction: Modification of Proteins Occurring in Isoelectric Focusing Electrophoresis and in Electrolytic Reactions in the Presence of High Salts

    PubMed Central

    2009-01-01

    Artifacts in two-dimensional electrophoresis (2-DE) caused by the presence of salts in isoelectric focusing (IEF) have been previously described as a result of increasing conductivity and inducing electroosmosis. However, electrolysis induced by the presence of salts should not be disregarded. In this study, electrolytic reduction−oxidation reaction (redox) was found to be enhanced in the presence of salts in IEF. The consequence of the electrolytic redox leads to acidification of the low-pH region and alkalization of the high-pH region within the immobilized pH gradient (IPG) strip. As a result, a breakdown of immobilized pH buffer near the high pH region of IPG strips along with reduction of basic proteins resulted in uncharacterized artifacts in 2-DE. Electrolytic reduction in the presence of alkali and alkaline metal ions was demonstrated to reduce 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), protein disulfide bonds, and protein carboxylic acids. Importantly, semipreparative electrolytic reduction of proteins can be carried out in the presence of sodium ions in a homemade electrolytic apparatus. These findings give additional explanations to the observed artifacts in 2-DE and reveal the unknown effects of salts in IEF. Moreover, we have provided a method with the potential to convert proteins or peptides to corresponding modified products containing aldehyde groups that can be used for conjugation with amine-containing compounds. PMID:19438264

  13. Triton X-114 cloud point extraction to subfractionate blood plasma proteins for two-dimensional gel electrophoresis.

    PubMed

    Jessen, Flemming; Wulff, Tune

    2015-09-15

    A simple and reproducible procedure for enrichment of a plasma protein subfraction suitable for two-dimensional polyacrylamide gel electrophoresis (2DE) was developed, using a Triton X-114-based cloud point extraction (CPE). Appropriate conditions for such a CPE procedure were found by SDS-PAGE to be a plasma protein concentration of about 10mg/ml in 3% (w/v) Triton X-114. 2DE of proteins obtained by CPE of 400 μl of human plasma revealed about 200 spots constituting a spot pattern very different from the pattern of total plasma. The CPE procedure only had a limited contribution to the technical variation. Identification of about 60 spots, representing only 22 proteins, revealed that several proteins in the obtained subfraction were present in more isoforms or modifications. Among these were apolipoproteins (A-1, D, E, L1, and M), haptoglobin-related protein, phosphatidylcholine-sterol acyltransferase, serum amyloid A, and serum paraoxonase/arylesterase 1, which are proteins of a hydrophobic nature, as in plasma they relate to lipoprotein particles. Thus, Triton X-114-based CPE is a simple plasma prefractionation tool, attractive for detailed 2DE studies of hydrophobic plasma proteins and their isoforms or modifications. PMID:26080275

  14. Extended length microchannels for high density high throughput electrophoresis systems

    DOEpatents

    Davidson, James C.; Balch, Joseph W.

    2000-01-01

    High throughput electrophoresis systems which provide extended well-to-read distances on smaller substrates, thus compacting the overall systems. The electrophoresis systems utilize a high density array of microchannels for electrophoresis analysis with extended read lengths. The microchannel geometry can be used individually or in conjunction to increase the effective length of a separation channel while minimally impacting the packing density of channels. One embodiment uses sinusoidal microchannels, while another embodiment uses plural microchannels interconnected by a via. The extended channel systems can be applied to virtually any type of channel confined chromatography.

  15. Micro-size polyacrylamide gel electrophoresis system

    NASA Astrophysics Data System (ADS)

    Hinson, W. G.; Pipkin, J. L.; Anson, J. F.; Casciano, D. A.; Burns, E. R.

    1987-09-01

    The development and characterization of a micro-size two-dimensional polyacrylamide gel electrophoresis system is described. Some of the techniques which have evolved with use of the system are also discussed. This apparatus has unique features which provide advantages over other small scale units. Up to ten first- and second-dimension gels can be processed simultaneously with excellent resolution of protein regions. Consistent reproducibility is possible from protein samples as small as 400 ng and individual protein regions as small as 1 pg can be visualized by silver staining of the two-dimensional gels. Similar sensitivities are achieved in autoradiographs of 3H-labeled proteins extracted from the nuclei of cultured cells. The application of this system in conjunction with flow cytometric examination of nuclear DNA and electrostatic cell sorting of specific cell nuclei to provide homogeneous sample populations, allows subtle variations in isotope incorporation in proteins to be detected; whereas many times in generalized tissue samples these changes are masked. Also, these techniques elucidate the effects of external stimuli (chemicals, drugs, or environment) on protein synthesis and phosphorylation for analyses and comparison. Fabrication drawings are available upon request.

  16. Microfab-less Microfluidic Capillary Electrophoresis Devices

    PubMed Central

    Segato, Thiago P.; Bhakta, Samir A.; Gordon, Matthew; Carrilho, Emanuel; Willis, Peter A.; Jiao, Hong; Garcia, Carlos D.

    2013-01-01

    Compared to conventional bench-top instruments, microfluidic devices possess advantageous characteristics including great portability potential, reduced analysis time (minutes), and relatively inexpensive production, putting them on the forefront of modern analytical chemistry. Fabrication of these devices, however, often involves polymeric materials with less-than-ideal surface properties, specific instrumentation, and cumbersome fabrication procedures. In order to overcome such drawbacks, a new hybrid platform is proposed. The platform is centered on the use of 5 interconnecting microfluidic components that serve as the injector or reservoirs. These plastic units are interconnected using standard capillary tubing, enabling in-channel detection by a wide variety of standard techniques, including capacitively-coupled contactless conductivity detection (C4D). Due to the minimum impact on the separation efficiency, the plastic microfluidic components used for the experiments discussed herein were fabricated using an inexpensive engraving tool and standard Plexiglas. The presented approach (named 52-platform) offers a previously unseen versatility: enabling the assembly of the platform within minutes using capillary tubing that differs in length, diameter, or material. The advantages of the proposed design are demonstrated by performing the analysis of inorganic cations by capillary electrophoresis on soil samples from the Atacama Desert. PMID:23585815

  17. Active airborne contamination control using electrophoresis

    SciTech Connect

    Veatch, B.D.

    1994-06-01

    In spite of our best efforts, radioactive airborne contamination continues to be a formidable problem at many of the Department of Energy (DOE) weapons complex sites. For workers that must enter areas with high levels of airborne contamination, personnel protective equipment (PPE) can become highly restrictive, greatly diminishing productivity. Rather than require even more restrictive PPE for personnel in some situations, the Rocky Flats Plant (RFP) is actively researching and developing methods to aggressively combat airborne contamination hazards using electrophoretic technology. With appropriate equipment, airborne particulates can be effectively removed and collected for disposal in one simple process. The equipment needed to implement electrophoresis is relatively inexpensive, highly reliable, and very compact. Once airborne contamination levels are reduced, less PPE is required and a significant cost savings may be realized through decreased waste and maximized productivity. Preliminary ``cold,`` or non-radioactive, testing results at the RFP have shown the technology to be effective on a reasonable scale, with several potential benefits and an abundance of applications.

  18. Simulating Electrophoresis with Discrete Charge and Drag

    NASA Astrophysics Data System (ADS)

    Mowitz, Aaron J.; Witten, Thomas A.

    A charged asymmetric rigid cluster of colloidal particles in saline solution can respond in exotic ways to an electric field: it may spin or move transversely. These distinctive motions arise from the drag force of the neutralizing countercharge surrounding the cluster. Because of this drag, calculating the motion of arbitrary asymmetric objects with nonuniform charge is impractical by conventional methods. Here we present a new method of simulating electrophoresis, in which we replace the continuous object and the surrounding countercharge with discrete point-draggers, called Stokeslets. The balance of forces imposes a linear, self-consistent relation among the drag and Coulomb forces on the Stokeslets, which allows us to easily determine the object's motion via matrix inversion. By explicitly enforcing charge+countercharge neutrality, the simulation recovers the distinctive features of electrophoretic motion to few-percent accuracy using as few as 1000 Stokeslets. In particular, for uniformly charged objects, we observe the characteristic Smoluchowski independence of mobility on object size and shape. We then discuss electrophoretic motion of asymmetric objects, where our simulation method is particularly advantageous. This work is supported by a Grant from the US-Israel Binational Science Foundation.

  19. Unexpected surface chemistry in capillaries for electrophoresis.

    PubMed

    Kaupp, S; Bubert, H; Baur, L; Nelson, G; Wätzig, H

    2000-10-13

    Good and reproducible capillary quality is needed to develop robust methods and to facilitate method transfer in CE. Physical surface defects no longer play a major role in variability of fused-silica capillaries. Nevertheless, problems are frequently being reported when buffers in the pH range between 4 and 7 are used. Thus the surface chemistry has been studied by X-ray photoelectron spectroscopy. Silicon-carbon bindings have been found on inner capillary surfaces for electrophoresis. This binding type is not completely removed by pre-conditioning with 1 M NaOH for 30 min. This corresponds to the result, that capillaries provide more stable migration times, especially in the pH range 4-7, when they are pre-conditioned for longer than 1 h. The origin of this Si-C bond is still not quite clear. They could be caused by graphite which is used during the fabrication of the raw cylinders prior to capillary drawing. Further investigations are intended in order to understand if there are any differences in surface carbon content from batch to batch and if this can influence experimental results in CE. A better understanding of the surface chemistry should not only improve robustness in CE, but also help to facilitate and accelerate capillary pre-conditioning and rinsing procedures to remove strongly adsorbed analytes or matrices. PMID:11100849

  20. Supported bilayer electrophoresis under controlled buffer conditions.

    PubMed

    Monson, Christopher F; Pace, Hudson P; Liu, Chunming; Cremer, Paul S

    2011-03-15

    A pH controlled flow cell device was constructed to allow electrophoretic movement of charged lipids and membrane associated proteins in supported phospholipid bilayers. The device isolated electrolysis products near the electrodes from the electrophoresis process within the bilayer. This allowed the pH over the bilayer region to remain within ±0.2 pH units or better over many hours at salt concentrations up to 10 mM. Using this setup, it was found that the electrophoretic mobility of a dye conjugated lipid (Texas Red 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (TR-DHPE)) was essentially constant between pH 3.3 and 9.3. In contrast, streptavidin, which was bound to biotinylated lipids, shifted from migrating cathodically at acidic pH values to migrating anodically under basic conditions. This shift was due to the modulation of the net charge on the protein, which changed the electrophoretic forces experienced by the macromolecule. The addition of a polyethylene glycol (PEG) cushion beneath the bilayer or the increase in the ionic strength of the buffer solution resulted in a decrease of the electroosmotic force experienced by the streptavidin with little effect on the Texas Red-DHPE. As such, it was possible in part to control the electrophoretic and electroosmotic contributions to streptavidin independently of one another. PMID:21319743

  1. Analysis of spiramycin by capillary electrophoresis.

    PubMed

    González-Hernández, R; Li, Y M; Van Schepdael, A; Roets, E; Hoogmartens, J

    1999-09-01

    The development and validation of an analytical method for the determination of spiramycin I in the presence of its related substances by capillary electrophoresis is shown. The separation, performed in a phosphate buffer (80 mM, pH 7.5) containing 12 mM cetyltrimethylammonium bromide (CTAB) and 20 mM sodium cholate, with a 50 microm ID and 44 cm long fused-silica capillary (36 cm effective length), applying a voltage of 12 kV (l approximately 80 microA), at 25 degrees C, is achieved in 15 min. Good selectivity among spiramycin I and its related substances was obtained. The influence of the buffer pH, and of the CTAB and sodium cholate concentrations was investigated. The method robustness, examined by means of a full-fraction factorial design, shows that it can be used within the limits set for the three parameters that were investigated. The method is linear (r = 0.9992) and precise (day-to-day corrected peak area repeatability, n = 18, relative standard deviation = 1.3%). The limits of detection and quantitation are 7 pg (0.025%) and 22 pg (0.08%), respectively, relative to a 2 mg/mL solution. PMID:10499332

  2. Capillary electrophoresis for total glycosaminoglycan analysis

    PubMed Central

    Ucakturk, Ebru; Cai, Chao; Li, Lingyun; Li, Guoyun; Zhang, Fuming; Linhardt, Robert J.

    2014-01-01

    A capillary zone electrophoresis-laser induced fluorescence detection (CZE-LIF) method was developed for the simultaneous analysis of disaccharides derived from heparan sulfate, chondroitin sulfate/dermatan sulfate, hyaluronan and keratan sulfate. Glycosaminoglycans (GAGs) were first depolymerized with the mixture of GAG lyases (heparinase I, II, III and chondroitinase ABC and chondroitinase AC II) and GAG endohydrolase (keratinase II) and the resulting disaccharides were derivatized by reductive amination with 2-aminoacridone. Nineteen fluorescently labeled disaccharides were separated using 50 mM phosphate buffer (pH 3.3) under reversed polarity at 25 kV. Using these conditions, all the disaccharides examined were baseline-separated in less then 25 min. This CZE-LIF method gave good reproducibility both migration time (≤ 1.03% for intra-day and ≤ 4.4% for inter-day) and the peak area values (≤ 5.6% for intra- and ≤ 8.69% for inter-day). This CZE-LIF method was used for profiling and quantification of GAG derivative disaccharides in bovine cornea. The results shows that the current CZE-LIF method offers fast, simple, sensitive, reproducible determination of disaccharides derived from total GAGs in a single run. PMID:24817364

  3. Contact charge electrophoresis: experiment and theory.

    PubMed

    Drews, Aaron M; Cartier, Charles A; Bishop, Kyle J M

    2015-04-01

    Contact charge electrophoresis (CCEP) uses steady electric fields to drive the continuous, oscillatory motion of conductive particles and droplets between two or more electrodes. These rapid oscillations can be rectified to direct the motion of objects within microfluidic environments using low-power, dc voltage. Here, we compare high precision experimental measurements of CCEP within a microfluidic system to equally detailed theoretical predictions on the motion of a conductive particle between parallel electrodes. We use a simple, capillary microfluidic platform that combines high-speed imaging with precision electrical measurements to enable the synchronized acquisition of both the particle location and the electric current due to particle motion. The experimental results are compared to those of a theoretical model, which relies on a Stokesian dynamics approach to accurately describe both the electrostatic and hydrodynamic problems governing particle motion. We find remarkable agreement between theory and experiment, suggesting that particle motion can be accurately captured by a combination of classical electrostatics and low-Reynolds number hydrodynamics. Building on this agreement, we offer new insight into the charge transfer process that occurs when the particle nears contact with an electrode surface. In particular, we find that the particle does not make mechanical contact with the electrode but rather that charge transfer occurs at finite surface separations of >0.1 μm by means of an electric discharge through a thin lubricating film. We discuss the implications of these findings on the charging of the particle and its subsequent dynamics. PMID:25785396

  4. Nonlinear electrophoresis of ideally polarizable particles

    NASA Astrophysics Data System (ADS)

    Figliuzzi, Bruno; Chan, Wai Hong Ronald; Buie, Cullen R.

    2013-11-01

    We focus in this presentation on the nonlinear electrophoresis of ideally polarizable particles. At high applied voltages, significant ionic exchanges occur between the EDL which surrounders the particle and the bulk solution. In this situation, the velocity field, the electric potential and the ionic concentration at the immediate vicinity of the particle are described by a complicated set of coupled nonlinear partial differential equations. These equations are classically considered in the limit of a weak applied field, which enables further analytical progress (Khair and Squires, Phys. Fluids, 2010). However, in the general case, the equation governing the electrophoretic motion of the particle must be solved numerically. In this study, we rely on a numerical approach to determine the electric potential, ionic concentration and velocity field in the bulk solution surrounding the particle. The numerical simulations use a pseudo-spectral which was used successfully by Chu and Bazant to determine the electric potential and the ionic concentration around an ideally polarizable metallic sphere (Physical Review E, 2006). Our numerical model also incorporates the steric model developed by Kilic et al. in 2007 to account for crowding effects in the electric double layer.

  5. Integrated polymerase chain reaction/electrophoresis instrument

    DOEpatents

    Andresen, Brian D.

    2000-01-01

    A new approach and instrument for field identification of micro-organisms and DNA fragments using a small and disposable device containing integrated polymerase chain reaction (PCR) enzymatic reaction wells, attached capillary electrophoresis (CE) channels, detectors, and read-out all on/in a small hand-held package. The analysis instrument may be made inexpensively, for example, of plastic, and thus is disposable, which minimizes cross contamination and the potential for false positive identification between samples. In addition, it is designed for multiple users with individual applications. The integrated PCR/CE is manufactured by the PCR well and CE channels are "stamped" into plastic depressions where conductive coatings are made in the wells and ends of the CE microchannels to carry voltage and current to heat the PCR reaction mixtures and simultaneously draw DNA bands up the CE channels. Light is transmitted through the instrument at appropriate points and detects PCR bands and identifies DNA fragments by size (retention time) and quantifies each by the amount of light generated as each phototransistor positioned below each CE channel detects a passing band. The instrument is so compact that at least 100 PCR/CE reactions/analyses can be performed easily on one detection device.

  6. Injector-concentrator electrodes for microchannel electrophoresis

    DOEpatents

    Swierkowski, Stefan P.

    2003-05-06

    An input port geometry, with injector-concentrator electrodes, for planar microchannel array for electrophoresis. This input port geometry enables efficient extraction and injection of the DNA sample from a single input port. The geometry, which utilizes injector-concentrator electrodes, allows simultaneous concentration, in different channels, of the sample into a longitudinally narrow strip just before releasing it for a run with enhanced injection spatial resolution, and time resolution. Optional multiple electrodes, at a different bias than the concentrator electrodes, may be used to discriminate against sample impurity ions. Electrode passivation can be utilized to prevent electrolysis. An additional electrode in or on the input hole can better define the initial loading. The injector-concentrator electrodes are positioned so that they cross the drift channel in a narrow strip at the bond plane between the top and bottom plates of the instrument and are located close to the inlet hole. The optional sample purification electrodes are located at a greater distance from the input hole than the injector-concentrate electrodes.

  7. Redox options in two-dimensional electrophoresis.

    PubMed

    Wait, R; Begum, S; Brambilla, D; Carabelli, A M; Conserva, F; Rocco Guerini, A; Eberini, I; Ballerio, R; Gemeiner, M; Miller, I; Gianazza, E

    2005-05-01

    Two-dimensional electrophoresis is usually run on fully reduced samples. Under these conditions even covalently bound oligomers are dissociated and individual polypeptide chains may be fully unfolded by both, urea and SDS, which maximizes the number of resolved components and allows their pI and M(r) to be most accurately evaluated. However, various electrophoretic protocols for protein structure investigation require a combination of steps under varying redox conditions. We review here some of the applications of these procedures. We also present some original data about a few related samples -- serum from four species: Homo sapiens, Mus musculus, Rattus norvegicus, Bos taurus -- which we run under fully unreduced and fully reduced conditions as well as with reduction between first and second dimension. We demonstrate that in many cases the unreduced proteins migrate with a better resolution than reduced proteins, mostly in the crowded 'alpha-globulin' area of pI 4.5-6 and M(r) 50-70 kDa. PMID:15744479

  8. Contactless conductivity detector for microchip capillary electrophoresis

    NASA Technical Reports Server (NTRS)

    Pumera, Martin; Wang, Joseph; Opekar, Frantisek; Jelinek, Ivan; Feldman, Jason; Lowe, Holger; Hardt, Steffen; Svehla, D. (Principal Investigator)

    2002-01-01

    A microfabricated electrophoresis chip with an integrated contactless conductivity detection system is described. The new contactless conductivity microchip detector is based on placing two planar sensing aluminum film electrodes on the outer side of a poly(methyl methacrylate) (PMMA) microchip (without contacting the solution) and measuring the impedance of the solution in the separation channel. The contactless route obviates problems (e.g., fouling, unwanted reactions) associated with the electrode-solution contact, offers isolation of the detection system from high separation fields, does not compromise the separation efficiency, and greatly simplifies the detector fabrication. Relevant experimental variables, such as the frequency and amplitude of the applied ac voltage or the separation voltage, were examined and optimized. The detector performance was illustrated by the separation of potassium, sodium, barium, and lithium cations and the chloride, sulfate, fluoride, acetate, and phosphate anions. The response was linear (over the 20 microM-7 mM range) and reproducible (RSD = 3.4-4.9%; n = 10), with detection limits of 2.8 and 6.4 microM (for potassium and chloride, respectively). The advantages associated with the contactless conductivity detection, along with the low cost of the integrated PMMA chip/detection system, should enhance the power and scope of microfluidic analytical devices.

  9. Fabricating PFPE Membranes for Capillary Electrophoresis

    NASA Technical Reports Server (NTRS)

    Lee, Michael C.; Willis, Peter A.; Greer, Frank; Rolland, Jason

    2009-01-01

    A process has been developed for fabricating perfluoropolyether (PFPE) membranes that contain microscopic holes of precise sizes at precise locations. The membranes are to be incorporated into laboratory-on-a-chip microfluidic devices to be used in performing capillary electrophoresis. The present process is a modified version of part of the process, described in the immediately preceding article, that includes a step in which a liquid PFPE layer is cured into solid (membrane) form by use of ultraviolet light. In the present process, one exploits the fact that by masking some locations to prevent exposure to ultraviolet light, one can prevent curing of the PFPE in those locations. The uncured PFPE can be washed away from those locations in the subsequent release and cleaning steps. Thus, holes are formed in the membrane in those locations. The most straightforward way to implement the modification is to use, during the ultraviolet-curing step, an ultraviolet photomask similar to the photomasks used in fabricating microelectronic devices. In lieu of such a photomask, one could use a mask made of any patternable ultraviolet-absorbing material (for example, an ink or a photoresist).

  10. Strongly nonlinear waves in capillary electrophoresis

    NASA Astrophysics Data System (ADS)

    Chen, Zhen; Ghosal, Sandip

    2012-05-01

    In capillary electrophoresis, sample ions migrate along a microcapillary filled with a background electrolyte under the influence of an applied electric field. If the sample concentration is sufficiently high, the electrical conductivity in the sample zone could differ significantly from the background. Under such conditions, the local migration velocity of sample ions becomes concentration-dependent, resulting in a nonlinear wave that exhibits shocklike features. If the nonlinearity is weak, the sample concentration profile, under certain simplifying assumptions, can be shown to obey Burgers’ equation [Ghosal and Chen, Bull. Math. Biol.BMTBAP0092-824010.1007/s11538-010-9527-2 72, 2047 (2010)], which has an exact analytical solution for arbitrary initial condition. In this paper, we use a numerical method to study the problem in the more general case where the sample concentration is not small in comparison to the concentration of background ions. In the case of low concentrations, the numerical results agree with the weakly nonlinear theory presented earlier, but at high concentrations, the wave evolves in a way that is qualitatively different.

  11. Robotics in biomedical chromatography and electrophoresis.

    PubMed

    Fouda, H G

    1989-08-11

    The ideal laboratory robot can be viewed as "an indefatigable assistant capable of working continuously for 24 h a day with constant efficiency". The development of a system approaching that promise requires considerable skill and time commitment, a thorough understanding of the capabilities and limitations of the robot and its specialized modules and an intimate knowledge of the functions to be automated. The robot need not emulate every manual step. Effective substitutes for difficult steps must be devised. The future of laboratory robots depends not only on technological advances in other fields, but also on the skill and creativity of chromatographers and other scientists. The robot has been applied to automate numerous biomedical chromatography and electrophoresis methods. The quality of its data can approach, and in some cases exceed, that of manual methods. Maintaining high data quality during continuous operation requires frequent maintenance and validation. Well designed robotic systems can yield substantial increase in the laboratory productivity without a corresponding increase in manpower. They can free skilled personnel from mundane tasks and can enhance the safety of the laboratory environment. The integration of robotics, chromatography systems and laboratory information management systems permits full automation and affords opportunities for unattended method development and for future incorporation of artificial intelligence techniques and the evolution of expert systems. Finally, humanoid attributes aside, robotic utilization in the laboratory should not be an end in itself. The robot is a useful tool that should be utilized only when it is prudent and cost-effective to do so. PMID:2671006

  12. Free zone electrophoresis simulation of static column electrophoresis in microgravity on shuttle flight STS-3

    NASA Technical Reports Server (NTRS)

    Todd, P. W.; Hjerten, S.

    1985-01-01

    Experiments were designed to replicate, as closely as possible in 1-G, the conditions of the STS-3 red blood cell (RBC) experiments. Free zone electrophoresis was the method of choice, since it minimizes the role of gravity in cell migration. The physical conditions of the STS-3 experiments were used, and human and rabbit RBC's fixed by the same method were the test particles. The effects of cell concentration, electroosmotic mobility, and sample composition were tested in order to seek explanations for the STS-3 results and to provide data on cell concentration effects for future zero-G separation on the continuous-flow zero-G electrophoretics separator.

  13. Determination of chemical concentration with a 2 dimensional CCD array in the Echelle grating spectrometer

    SciTech Connect

    Lewis, D.K.; Stevens, C.G.

    1994-11-15

    The Echelle grating spectrometer (EGS) uses a stepped Echelle grating, prisms and a folded light path to miniaturize an infrared spectrometer. Light enters the system through a slit and is spread out along Y by a prism. This light then strikes the grating and is diffracted out along X. This spreading results in a superposition of spectral orders since the grating has a high spectral range. These orders are then separated by again passing through a prism. The end result of a measurement is a 2 dimensional image which contains the folded spectrum of the region under investigation. The data lies in bands from top to bottom, for example, with wavenumber increments as small as 0.1 lying from left to right such that the right end of band N is the same as the left end of band N+1. This is the image which must be analyzed.

  14. Time-resolved spatial phase measurements with 2-dimensional spectral interferometry

    NASA Astrophysics Data System (ADS)

    Childress, Colby; Planchon, Thomas; Amir, Wafa; Squier, Jeff A.; Durfee, Charles G.

    2007-03-01

    We are using 2-dimensional spectral interferometry for sensitive measurements of spatial phase distortions. The reference pulse and the time-delayed probe pulse are coincident on an imaging spectrometer, yielding spectral and spatial phase information. This technique offers the potential of higher sensitivity than traditional spatial interferometry since there are many fringes of data for each spatial point. We illustrate this technique with measurements of the thermal lensing profile in a cryogenically cooled Ti:sapphire amplifier crystal that is pumped by tens of watts of power from four frequency-doubled Nd:YLF lasers running at 1 kHz. By adjusting the relative delay of the probe and reference pulses, we characterize the thermal transients during and after the pump pulses. We compare the measured transient thermal profiles with those calculated with a finite-element model.

  15. The structural identification of a methyl analog of methaqualone via 2-dimensional NMR techniques.

    PubMed

    Angelos, S A; Lankin, D C; Meyers, J A; Raney, J K

    1993-03-01

    A submission to the Drug Enforcement Administration North Central Laboratory of a substance believed to be a structural analog of methaqualone hydrochloride precipitated an interest in being able to obtain a rapid and positive identification of such compounds. Both mass spectrometry and proton NMR spectroscopy (1-dimensional) provided evidence to suggest that the structural analog possessed a second methyl group in the molecule, relative to methaqualone, and that the methyl group was attached to the existing methyl-substituted phenyl ring. By application of proton 2-dimensional (2-D) NMR techniques, specifically the homonuclear shift correlation spectroscopy (COSY) and 2-D NOE (NOESY), the precise location of the methyl group in this unknown methaqualone analog was established and shown to have the structure 2. PMID:8455002

  16. 2-dimensional simulations of electrically asymmetric capacitively coupled RF-discharges

    NASA Astrophysics Data System (ADS)

    Mohr, Sebastian; Schulze, Julian; Schuengel, Edmund; Czarnetzki, Uwe

    2011-10-01

    Capactively coupled RF-discharges are widely used for surface treatment like the deposition of thin films. For industrial applications, the independent control of the ion flux to and the mean energy of the electrons impinging on the surfaces is desired. Experiments and 1D3v-PIC/MCC-simulations have shown that this independent control is possible by applying a fundamental frequency and its second harmonic to the powered electrode. This way, even in geometrically symmetric discharges, as they are often used in industrial reactors, a discharge asymmetry can be induced electrically, hence the name Electrical Asymmetry Effect (EAE). We performed 2D-simulations of electrically asymmetric discharges using HPEM by the group of Mark Kushner, a simulation tool suitable for simulating industrial reactors. First results are presented and compared to previously obtained experimental and simulation data. The comparison shows that for the first time, we succeeded in simulating electrically asymmetric discharges with a 2-dimensional simulation. Capactively coupled RF-discharges are widely used for surface treatment like the deposition of thin films. For industrial applications, the independent control of the ion flux to and the mean energy of the electrons impinging on the surfaces is desired. Experiments and 1D3v-PIC/MCC-simulations have shown that this independent control is possible by applying a fundamental frequency and its second harmonic to the powered electrode. This way, even in geometrically symmetric discharges, as they are often used in industrial reactors, a discharge asymmetry can be induced electrically, hence the name Electrical Asymmetry Effect (EAE). We performed 2D-simulations of electrically asymmetric discharges using HPEM by the group of Mark Kushner, a simulation tool suitable for simulating industrial reactors. First results are presented and compared to previously obtained experimental and simulation data. The comparison shows that for the first time, we

  17. A New Electrophoresis Technique to Seperate Microsatellite Alleles

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditional agarose and polyacrylamide gel electrophoresis have been used commonly for microsatellite (simple sequence repeats, SSRs) analysis, but they are labor- intensive and not always able to provide accurate sizes for different alleles. Capillary sequencers provide automated analysis and accur...

  18. Microchannel DNA Sequencing by End-Labelled Free Solution Electrophoresis

    SciTech Connect

    Barron, A.

    2005-09-29

    The further development of End-Labeled Free-Solution Electrophoresis will greatly simplify DNA separation and sequencing on microfluidic devices. The development and optimization of drag-tags is critical to the success of this research.

  19. Integration of amperometric sensors for microchip capillary electrophoresis application

    NASA Astrophysics Data System (ADS)

    Dicorato, F.; Moore, E.; Glennon, J.

    2011-08-01

    Capillary electrophoresis is a technique for the separation and analysis of chemical compounds. Techniques adopted from the microchip technology knowledge have led to recent developments of electrophoresis system with integration on microchip. Microchip Capillary Electrophoresis (μCE) systems offer a series of advantages as easy integration for Lab-on-a-chip applications, high performance, portability, speed, minimal solvent and sample requirements. A new technological challenge aims at the development of an economic modular microchip capillary electrophoresis systems using separable and independent units concerning the sensor. In this project we worked on the development of an interchangeable amperometric sensor in order to provide a solution to such electrode passivation and facilitating the use of tailored sensors for specific analyte detection besides. Fluidic chips have been machined from cyclic olefin polymer pallets (Zeonor®) using a micro-injection molding machine.

  20. CAPILLARY ELECTROPHORESIS FOR THE CHARACTERIZATION OF HUMIC SUBSTANCES

    EPA Science Inventory

    The potential of high performance capillary electrophoresis (HPCE), especially in the free solution mode (FSCE), is demonstrated for the analysis/characterization of environmental humic substances (HUS). he very high efficiency of HPCE separations allows the production of electro...

  1. Nonlinear electrophoresis of ideally polarizable particles

    NASA Astrophysics Data System (ADS)

    Figliuzzi, B.; Chan, W. H. R.; Moran, J. L.; Buie, C. R.

    2014-10-01

    We focus in this paper on the nonlinear electrophoresis of ideally polarizable particles. At high applied voltages, significant ionic exchange occurs between the electric double layer, which surrounds the particle, and the bulk solution. In addition, steric effects due to the finite size of ions drastically modify the electric potential distribution in the electric double layer. In this situation, the velocity field, the electric potential, and the ionic concentration in the immediate vicinity of the particle are described by a complicated set of coupled nonlinear partial differential equations. In the general case, these equations must be solved numerically. In this study, we rely on a numerical approach to determine the electric potential, the ionic concentration, and the velocity field in the bulk solution surrounding the particle. The numerical simulations rely on a pseudo-spectral method which was used successfully by Chu and Bazant [J. Colloid Interface Sci. 315(1), 319-329 (2007)] to determine the electric potential and the ionic concentration around an ideally polarizable metallic sphere. Our numerical simulations also incorporate the steric model developed by Kilic et al. [Phys. Rev. E 75, 021502 (2007)] to account for crowding effects in the electric double layer, advective transport, and for the presence of a body force in the bulk electrolyte. The simulations demonstrate that surface conduction significantly decreases the electrophoretic mobility of polarizable particles at high zeta potential and at high applied electric field. Advective transport in the electric double layer and in the bulk solution is also shown to significantly impact surface conduction.

  2. Band broadening of DNA fragments isolated by polyacrylamide gel electrophoresis in capillary electrophoresis.

    PubMed

    Kaneta, Takashi; Ogura, Takehito; Yamato, Shuhei; Imasaka, Totaro

    2012-02-01

    Polyacrylamide gel electrophoresis (PAGE) is used frequently for isolation and purification of DNA fragments. In the present study, DNA fragments extracted from polyacrylamide gels showed significant band broadening in capillary electrophoresis (CE). A pHY300PLK (a shuttle vector functioning in Escherichia coli and Bacillus subtilis) marker, which contained nine fragments ranging from 80 to 4870 bp, was separated by PAGE, and each fragment was isolated by phenol/chloroform extraction and ethanol precipitation. After extraction from the polyacrylamide gel, the peaks of the isolated DNA fragments exhibited band broadening in CE, where a linear poly(ethylene oxide) was used as a sieving matrix. The theoretical plate numbers of the DNA fragments contained in the pHY300PLK marker were >10(6) for all the fragments before extraction. However, the DNA fragments extracted from the polyacrylamide gel showed decreased theoretical plate numbers (5-20 times smaller). The degradation of the theoretical plate number was significant for middle sizes of the DNA fragments ranging from 489 to 1360 bp, whereas the largest and smallest fragments (80 and 4870 bp) had no obvious influence. The band broadening was attributed to contamination of the DNA fragments by polyacrylamide fibers during the separation and extraction process. PMID:22258810

  3. Novel separation and detection methods of DNA fragments in electrophoresis

    SciTech Connect

    Chan, King Cheung

    1993-01-27

    A charge-coupled device (CCD) based electrophoresis system was developed. The system allowed non-destructive, sensitive, and on-line detection of native DNA in slab-gel electrophoresis via ultraviolet absorption measurement. The detection limit of double-stranded DNA fragment was 5 ng per band. Since the amount of DNA used in this experiment was typical, the CCD-based system could be readily implemented in molecular biology. Gel-filled and non-gel sieving capillary electrophoresis was developed for rapid and efficient separation of double-stranded DNA fragments. For the gel-filled CE separation a new gel matrix, the HydroLink gel (HL), was used. The HL capillary gel was easier to cast than the polyacrylamide capillary gel. For the non-gel separation, a GC capillary was used as the separation chamber, and cellulose additive was included in the electrophoresis as the sieving medium. Indirect fluorometry was applied in non-gel and gel electrophoresis for the detection of DNA fragments. This method allowed non-destructive and on-line detection of DNA during electrophoresis. The amount of DNA used with this method was comparable to those obtained with absorption measurement.

  4. Novel separation and detection methods of DNA fragments in electrophoresis

    SciTech Connect

    Chan, K.C.

    1992-01-01

    A charge-coupled device (CCD) based electrophoresis system was developed. The system allowed non-destructive, sensitive, and on-line detection of native DNA in slab-gel electrophoresis via ultraviolet absorption measurement. The detection limit of double-stranded DNA fragment was 5 ng per band. Since the amount of DNA used in this experiment was typical, the CCD-based system could be readily implemented in molecular biology. Gel-filled and non-gel sieving capillary electrophoresis (CE) was developed for rapid and efficient separation of double-stranded DNA fragments. For the gel-filled CE separation a new gel matrix, the HydroLink gel (HL), was used. The HL capillary gel was easier to cast than the polyacrylamide capillary gel. For the non-gel separation, a GC capillary was used as the separation chamber, and cellulose additive was included in the electrophoresis as the sieving medium. Indirect fluorometry was applied in non-gel and gel electrophoresis for the detection of DNA fragments. This method allowed nondestructive and on-line detection of DNA during electrophoresis. The amount of DNA used with this method was comparable to those obtained with absorption measurement.

  5. Characterization of copolymer latexes by capillary electrophoresis.

    PubMed

    Anik, Nadia; Airiau, Marc; Labeau, Marie-Pierre; Bzducha, Wojciech; Cottet, Hervé

    2010-02-01

    Latexes are widely used for industrial applications, including decorative paints, binders for the papermaking industry, and drilling fluids for oil-field applications. In this work, the interest of capillary zone electrophoresis (CE) for the characterization of hydrophobic block copolymer latexes obtained by the conventional emulsion polymerization technique consisting of a core of polystyrene (PS) surrounded by a layer of poly(ethyl acrylate) (PEA) has been investigated. The PEA part of the copolymer can be partially hydrolyzed in poly(acrylic acid) (PAA) leading to PS-PEA-AA water-soluble amphiphilic copolymer having high viscosifying properties. The main purpose of this work was to evaluate the potential of CE for the characterization of the latexes at the different stages of the synthesis (PS core, PS-PEA diblock latex, and hydrolyzed PS-PEA-AA gel). The main analytical issues were to state (i) if there was free PS or PEA homopolymer latexes in the PS-PEA latex sample and (ii) if there was free PS, PEA, PS-PEA latexes, or free PAA chains in the PS-PEA-AA gel. Within this scope, this work describes the optimization of the selectivity of the separation between the different species (PS, PEA particles in the not hydrolyzed diblock latex and PS, PEA, PS-PEA particles as well as the polymer PAA chains in the PS-PEA-AA diblock gel sample obtained by latter latex hydrolysis). For that purpose, several experimental parameters were investigated such as pH and ionic strength of the background electrolyte (BGE) or the concentration of neutral surfactant added in the BGE. A challenging issue was to overcome the high viscosity of the PS-PEA-AA gel. This was resolved by the addition of 10 mM neutral surfactant in the gel sample and in the BGE. Finally, it is demonstrated that, within the detection limits, CE is a suitable analytical tool for controlling and monitoring the syntheses of these latexes and for intrinsically characterizing the distribution in charge density of

  6. Differentiation between fresh and frozen-thawed sea bass (Dicentrarchus labrax) fillets using two-dimensional gel electrophoresis.

    PubMed

    Ethuin, Pierrette; Marlard, Sylvain; Delosière, Mylène; Carapito, Christine; Delalande, François; Van Dorsselaer, Alain; Dehaut, Alexandre; Lencel, Valérie; Duflos, Guillaume; Grard, Thierry

    2015-06-01

    This study aimed to identify a protein marker that can differentiate between fresh skinless and frozen-thawed sea bass (Dicentrarchus labrax) fillets using the two-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Distinct gel patterns, due to proteins with low molecular weight and low isoelectric points, distinguished fresh fillets from frozen-thawed ones. Frozen-thawed fillets showed two specific protein spots as early as the first day of the study. However, these spots were not observed in fresh fillets until at least 13days of storage between 0 and 4°C, fillets were judged, beyond this period, fish were unfit for human consumption as revealed by complementary studies on fish spoilage indicators namely total volatile basic nitrogen and biogenic amines. Mass spectrometry identified the specific proteins as parvalbumin isoforms. Parvalbumins may thus be useful markers of differentiation between fresh and frozen-thawed sea bass fillets. PMID:25624236

  7. Recent Developments in Instrumentation for Capillary Electrophoresis and Microchip-Capillary Electrophoresis

    PubMed Central

    Felhofer, Jessica L.; Blanes, Lucas; Garcia, Carlos D.

    2010-01-01

    Over the last years there has been an explosion in the number of developments and applications of capillary electrophoresis (CE) and microchip-CE. In part, this growth has been the direct consequence of recent developments in instrumentation associated with CE. This review, which is focused on contributions published in the last five years, is intended to complement the papers presented in this special issue dedicated to Instrumentation and to provide an overview on the general trend and some of the most remarkable developments published in the areas of high voltage power supplies, detectors, auxiliary components, and compact systems. It also includes few examples of alternative uses of and modifications to traditional CE instruments. PMID:20665910

  8. Nonlinear Exact Solutions of the 2-Dimensional Rotational Euler Equations for the Incompressible Fluid

    NASA Astrophysics Data System (ADS)

    An, Hong-Li; Yang, Jin-Jing; Yuen, Man-Wai

    2015-05-01

    In this paper, the Clarkson-Kruskal direct approach is employed to investigate the exact solutions of the 2-dimensional rotational Euler equations for the incompressible fluid. The application of the method leads to a system of completely solvable ordinary differential equations. Several special cases are discussed and novel nonlinear exact solutions with respect to variables x and y are obtained. It is of interest to notice that the pressure p is obtained by the second kind of curvilinear integral and the coefficients of the nonlinear solutions are solitary wave type functions like tanh(kt/2) and sech (kt/2) due to the rotational parameter k ≠ 0. Such phenomenon never appear in the classical Euler equations wherein the Coriolis force arising from the gravity and Earth's rotation is ignored. Finally, illustrative numerical figures are attached to show the behaviors that the exact solutions may exhibit. Supported by the National Natural Science Foundation of China under Grant No. 11301269, Jiangsu Provincial Natural Science Foundation of China under Grant No. BK20130665, the Fundamental Research Funds KJ2013036 for the Central Universities, Student Research Training under Grant No. 1423A02 of Nanjing Agricultural University, and the Research Grant RG21/2013-2014R from the Hong Kong Institute of Education

  9. Role of surface defects on the formation of the 2-dimensional electron gas at polar interfaces

    NASA Astrophysics Data System (ADS)

    Artacho, Emilio; Aguado-Puente, Pablo

    2014-03-01

    The discovery of a 2-dimensional electron gas (2DEG) at the interface between two insulators, LaAlO3 and SrTiO3, has fuelled a great research activity on this and similar systems in the last years. The electronic reconstruction model, typically invoked to explain the formation of the 2DEG, while being intuitive and successful on predicting fundamental aspects of this phenomenon like the critical thickness of LaAlO3, fails to explain many other experimental observations. Oxygen vacancies, on the other hand, are known to dramatically affect the physical behaviour of this system, but their role at the atomic level is far from well understood. Here we perform ab initio simulations in order to assess whether the formation of oxygen vacancies at the surface of the polar material can account for various recent experimental results that defy the current theoretical understanding of these interfaces. We simulate SrTiO3/LaAlO3 slabs with various concentrations of surface oxygen vacancies and analyze the role of the defects on the formation of the metallic interface, their electrostatic coupling with the 2DEG and the interplay with the different instabilities of the materials involved. Financial support from Spanish MINECO under grant FIS2012-37549-C05-01. Computational resources provided by the Red Espñola de Supercomputación and DIPC.

  10. A 2-dimensional fully analytical model for design of high voltage junction barrier Schottky (JBS) diodes

    NASA Astrophysics Data System (ADS)

    Radhakrishnan, Rahul; Zhao, Jian H.

    2011-09-01

    A physics-based closed form analytical model for the reverse leakage current of a high voltage junction barrier Schottky (JBS) diode is developed and shown to agree with experimental results. Maximum electric field "seen" by the Schottky contact is calculated from first principles by a 2-dimensional method as a function of JBS diode design parameters and confirmed by numerical simulations. Considering thermionic emission under image force barrier lowering and quantum mechanical tunneling, electric field at the Schottky contact is then related to reverse current. In combination with previously reported forward current and resistance models, this gives a complete I- V relationship for the JBS diode. A layout of interdigitated stripes of P-N and Schottky contacts at the anode is compared theoretically with a honeycomb layout and the 2-D model is extended to the 3-D honeycomb structure. Although simulation and experimental results from 4H-Silicon Carbide (SiC) diodes are used to validate it, the model itself is applicable to all JBS diodes.

  11. A 2-dimensional MHD code & survey of the ``buckling'' phenomenon in cylindrical magnetic flux compression experiments

    NASA Astrophysics Data System (ADS)

    Xiao, Bo; Wang, Ganghua; Gu, Zhuowei; Computational Physics Team

    2015-11-01

    We made a 2-dimensional magneto-hydrodynamics Lagrangian code. The code handles two kinds of magnetic configuration, a (x-y) plane with z-direction magnetic field Bz and a (r-z) plane with θ-direction magnetic field Bθ. The solving of the MHD equations is split into a pure dynamical step (i.e., ideal MHD) and a diffusion step. In the diffusion step, the Joule heat is calculated with a numerical scheme based on an specific form of the Joule heat production equation, ∂eJ/∂t = ∇ . (η/μ0 º × (∇ × º)) -∂/∂t (1/2μ0 B2) , where the term ∂/∂t (1/2μ0 B2) is the magnetic field energy variation caused solely by diffusion. This scheme insures the equality of the total Joule heat produced and the total electromagnetic energy lost in the system. Material elastoplasticity is considered in the code. An external circuit is coupled to the magneto-hydrodynamics and a detonation module is also added to enhance the code's ability for simulating magnetically-driven compression experiments. As a first application, the code was utilized to simulate a cylindrical magnetic flux compression experiment. The origin of the ``buckling'' phenomenon observed in the experiment is explored.

  12. Electronic thermal conductivity of 2-dimensional circular-pore metallic nanoporous materials

    NASA Astrophysics Data System (ADS)

    Huang, Cong-Liang; Lin, Zi-Zhen; Luo, Dan-Chen; Huang, Zun

    2016-09-01

    The electronic thermal conductivity (ETC) of 2-dimensional circular-pore metallic nanoporous material (MNM) was studied here for its possible applications in thermal cloaks. A simulation method based on the free-electron-gas model was applied here without considering the quantum effects. For the MNM with circular nanopores, there is an appropriate nanopore size for thermal conductivity tuning, while a linear relationship exists for this size between the ETC and the porosity. The appropriate nanopore diameter size will be about one times that of the electron mean free path. The ETC difference along different directions would be less than 10%, which is valuable when estimating possible errors, because the nanoscale-material direction could not be controlled during its application. Like nanoparticles, the ETC increases with increasing pore size (diameter for nanoparticles) while the porosity was fixed, until the pore size reaches about four times that of electron mean free path, at which point the ETC plateaus. The specular coefficient on the surface will significantly impact the ETC, especially for a high-porosity MNM. The ETC can be decreased by 30% with a tuning specular coefficient.

  13. Data assimilation technique of 2-dimensional vertical temperature transport model (Case study: Tropical Pacific Ocean)

    NASA Astrophysics Data System (ADS)

    Nurfitri, Suliskania; Putri, Mutiara Rachmat

    2015-09-01

    Data assimilation technique was applied into 2-dimensional vertical temperature transport model to improve temperature results especially at thermocline layer that have large change toward depth. The simple case experiment only applies on baroclinic condition in Tropical Pacific Ocean (2°N and 137°E - 140°W). Model simulation was running for 2 years from January 1st 2011 to December 31st 2012 from surface to 500 m depth and verified to observation data from TAO (Tropical Atmosphere Ocean) at 3 locations, 147°E, 165°E, and 170°W. Data assimilation procedure which applied at 3 locations (156°E, 180°W, and 155°W) using Cressman analysis technique can reduce model's RMSE (Root Mean Square Error) toward depth until 55,7% and toward time until 64,5%. The largest error of model was found at 200 m depth, while the smallest found at the surface and 500 m depth.

  14. Nonlinear electrophoresis for purification of soil DNA for metagenomics.

    PubMed

    Engel, Katja; Pinnell, Lee; Cheng, Jiujun; Charles, Trevor C; Neufeld, Josh D

    2012-01-01

    Purification of microbial DNA from soil is challenging due to the co-extraction of humic acids and associated phenolic compounds that inhibit subsequent cloning, amplification or sequencing. Removal of these contaminants is critical for the success of metagenomic library construction and high-throughput sequencing of extracted DNA. Using three different composite soil samples, we compared a novel DNA purification technique using nonlinear electrophoresis on the synchronous coefficient of drag alteration (SCODA) instrument with alternate purification methods such as direct current (DC) agarose gel electrophoresis followed by gel filtration or anion exchange chromatography, Wizard DNA Clean-Up System, and the PowerSoil DNA Isolation kit. Both nonlinear and DC electrophoresis were effective at retrieving high-molecular weight DNA with high purity, suitable for construction of large-insert libraries. The PowerSoil DNA Isolation kit and the nonlinear electrophoresis had high recovery of high purity DNA suitable for sequencing purposes. All methods demonstrated high consistency in the bacterial community profiles generated from the DNA extracts. Nonlinear electrophoresis using the SCODA instrument was the ideal methodology for the preparation of soil DNA samples suitable for both high-throughput sequencing and large-insert cloning applications. PMID:22056233

  15. Electrophoresis of DNA and other polyelectrolytes: Physical mechanisms

    NASA Astrophysics Data System (ADS)

    Viovy, Jean-Louis

    2000-07-01

    The dramatic recent advances in molecular biology, which have opened a new era in medicine and biotechnology, rely on improved techniques to study large molecules. Electrophoresis is one of the most important of these. Separation of DNA by size, in particular, is at the heart of genome mapping and sequencing and is likely to play an increasing role in diagnosis. This article reviews, from the point of view of a physicist, the mechanisms responsible for electrophoretic separation of polyelectrolytes. This separation is mainly performed in gels, and a wide variety of migration mechanisms can come into play, depending on the polyelectrolyte's architecture, on the electric fields applied, and on the properties of the gel. After a brief review of the thermodynamic and electrohydrodynamic principles relating to polyelectrolyte solutions, the author treats the phenomenology of electrophoresis and describes the conceptual and theoretical tools in the field. The reptation mechanisms, by which large flexible polyelectrolytes thread their way through the pores of the gel matrix, play a prominent role. Biased reptation, the extension of this model to electrophoresis, provides a very intuitive framework within which numerous physical ideas can be introduced and discussed. It has been the most popular theory in this domain, and it remains an inspiring concept for current development. There have also been important advances in experimental techniques such as single-molecule viodeomicroscopy and the development of nongel separation media and mechanisms. These, in turn, form the basis for fast-developing and innovative technologies like capillary electrophoresis, electrophoresis on microchips, and molecular ratchets.

  16. 2-DE combined with two-layer feature selection accurately establishes the origin of oolong tea.

    PubMed

    Chien, Han-Ju; Chu, Yen-Wei; Chen, Chi-Wei; Juang, Yu-Min; Chien, Min-Wei; Liu, Chih-Wei; Wu, Chia-Chang; Tzen, Jason T C; Lai, Chien-Chen

    2016-11-15

    Taiwan is known for its high quality oolong tea. Because of high consumer demand, some tea manufactures mix lower quality leaves with genuine Taiwan oolong tea in order to increase profits. Robust scientific methods are, therefore, needed to verify the origin and quality of tea leaves. In this study, we investigated whether two-dimensional gel electrophoresis (2-DE) and nanoscale liquid chromatography/tandem mass spectroscopy (nano-LC/MS/MS) coupled with a two-layer feature selection mechanism comprising information gain attribute evaluation (IGAE) and support vector machine feature selection (SVM-FS) are useful in identifying characteristic proteins that can be used as markers of the original source of oolong tea. Samples in this study included oolong tea leaves from 23 different sources. We found that our method had an accuracy of 95.5% in correctly identifying the origin of the leaves. Overall, our method is a novel approach for determining the origin of oolong tea leaves. PMID:27283647

  17. Study of capillary electrophoresis chip with conductivity detection

    NASA Astrophysics Data System (ADS)

    Wei, Wang; Li, Tian; Liu, Xiao-wei; Lei, Xuan

    2006-04-01

    Microchip electrophoresis technique have been developed fully these years, especially the detection methods have been the research focuses. With the help of MEMS technique, the system integrating function are more and more. This paper presented a novel design and fabrication method about this electrophoresis chip with the detection electrodes. The two-layer structure, one is PDMS substrate to fabricate the electrophoresis microchannel, the other is the glass substrate with the Pt electrodes, are bonded together. The soft lithography with the two molds is developed to accommodate the PDMS curing and peeling off. Then the inorganic cation Cu 2+ is introduced and tested the project feasibility. All these works are laid a stable foundation for the chip appliance.

  18. THERMAL DETECTION OF DNA AND PROTEINS DURING GEL ELECTROPHORESIS

    SciTech Connect

    R. JOHNSTON

    2000-08-01

    This is the final report of a three-year, Laboratory-Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). The goal of this project was to try to detect unstained, untagged, unlabeled DNA bands in real-time during gel electrophoresis using simple thermal measurements. The technical and ES&H advantages to this approach could potentially be quite significant, especially given the extreme importance of gel electrophoresis to a wide variety of practical and research fields. The project was unable to demonstrate sufficient thermal sensitivity to detect DNA bands. It is clear that we still do not understand the gel electrophoresis phenomenon very well. The temperature control techniques developed during the course of this project have other useful applications.

  19. Nonlinear electrophoresis in the presence of dielectric decrement.

    PubMed

    Figliuzzi, B; Chan, W H R; Buie, C R; Moran, J L

    2016-08-01

    The nonlinear phenomena that occur in the electric double layer (EDL) that forms at charged surfaces strongly influence electrokinetic effects, including electro-osmosis and electrophoresis. In particular, saturation effects due to either dielectric decrement or ion crowding effects are of paramount importance. Dielectric decrement significantly influences the ionic concentration in the EDL at high ζ potential, leading to the formation of a condensed layer near the particle's surface. In this article, we present a model incorporating both steric effects due to the finite size of ions and dielectric decrement to describe the physics in the electric double layer. The model remains valid in both weakly and strongly nonlinear regimes, as long as the electric double layer remains in quasiequilibrium. We apply this model to the study of two archetypal problems in electrokinetics, namely the electrophoresis of particles with fixed surface charges and the electrophoresis of ideally polarizable particles. PMID:27627400

  20. Enantiomeric resolution of multiple chiral centres racemates by capillary electrophoresis.

    PubMed

    Ali, Imran; Suhail, Mohd; Al-Othman, Zeid A; Alwarthan, Abdulrahman; Aboul-Enein, Hassan Y

    2016-05-01

    Enantiomeric resolution of multichiral centre racemates is an important area as some multichiral centre racemates are of great medicinal importance. However, enantioseparation of such types of racemates is a challenging task. Amongst many analytical techniques, capillary electrophoresis is a powerful technique and may be used to resolve such racemates. Only few papers are available describing enantiomeric resolution of such racemates. Therefore, efforts have been made to describe the enantiomeric resolution of multichiral centre racemates by capillary electrophoresis. This article discusses the importance of multichiral racemates, the need for capillary electrophoresis in enantiomeric resolution and chiral resolution of multichiral centre racemates using various chiral selectors. Further, attempts have been made to discuss the future challenges and prospects of enantiomeric resolution of multichiral racemates. The various chiral selectors used for the purpose are chiral crown ether, cyclodextrins, polysaccharides, macrocyclic glycopeptide antibiotics and ligand exchange. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26840015

  1. Inexpensive and safe DNA gel electrophoresis using household materials.

    PubMed

    Ens, S; Olson, A B; Dudley, C; Ross, N D; Siddiqi, A A; Umoh, K M; Schneegurt, M A

    2012-01-01

    Gel electrophoresis is the single most important molecular biology technique and it is central to life sciences research, but it is often too expensive for the secondary science classroom or homeschoolers. A simple safe low-cost procedure is described here that uses household materials to construct and run DNA gel electrophoresis. Plastic containers are fitted with aluminum foil electrodes and 9-V batteries to run food-grade agar-agar gels using aquarium pH buffers and then stained with gentian violet. This activity was tested in a high school biology classroom with significantly positive responses on postactivity reflective surveys. The electrophoresis activity addresses several Life Science Content Standard C criteria, including aspects of cell biology, genetics, and evolution. It also can be used to teach aspects of motion and force in the physical science classroom. PMID:22615228

  2. Micro free-flow electrophoresis: theory and applications

    PubMed Central

    Turgeon, Ryan T.

    2009-01-01

    Free-flow electrophoresis (FFE) is a technique that performs an electrophoretic separation on a continuous stream of analyte as it flows through a planar flow channel. The electric field is applied perpendicularly to the flow to deflect analytes laterally according to their mobility as they flow through the separation channel. Miniaturization of FFE (μFFE) over the past 15 years has allowed analytical and preparative separation of small volume samples. Advances in chip design have improved separations by reducing interference from bubbles generated by electrolysis. Mechanisms of band broadening have been examined theoretically and experimentally to improve resolution in μFFE. Separations using various modes such as zone electrophoresis, isoelectric focusing, isotachophoresis, and field-step electrophoresis have been demonstrated. PMID:19290514

  3. Capillary and microchip electrophoresis: challenging the common conceptions.

    PubMed

    Breadmore, Michael C

    2012-01-20

    Capillary electrophoresis (CE) has long been regarded as a powerful analytical separation technique that is an alternative to more traditional methods such as gel electrophoresis (GE) and liquid chromatography (LC). It is often touted as having a number of advantages over both of these, such as speed, flexibility, portability, sample and reagent requirements and cost, but also a number of disadvantages such as reproducibility and sensitivity. Microchip electrophoresis (ME), the next evolutionary step, miniaturised CE further providing improvements in speed and sample requirements as well as the possibility to perform more complex and highly integrated analyses. CE and ME are seen as a viable alternative to GE, but are often considered to be inferior to LC. This review will consider the strengths and weaknesses of both CE and ME and will challenge the common conceptions held about these. PMID:22000781

  4. Preparative cell electrophoresis at 1 and 0 gravity

    NASA Technical Reports Server (NTRS)

    Van Oss, C. J.; Bronson, P. M.

    1979-01-01

    It is attempted to show that the use of heavy water (D2O) as starting cushion for the cells combines the advantages of the required density difference, with no lasting biochemical or physiochemical influence on the cells. Phosphate buffers of low ionic strength were prepared in distilled water or heavy water. A vertical starch gel electrophoresis was used to support a cylindrical polystyrene electrophoresis tube used for lymphocyte separations, 25 cm in length and 0.75 cm I.D., prepared from a 10 ml disposable pipet. Erythrocyte separations were carried out in a jacketed rectangular plexiglas chamber. It is pointed out that the described preparative D2O gradient electrophoresis method cannot be readily used for the measurement of electrophoretic mobilities for analytical purposes. However, for the preparative separation of cells with only slightly different electrokinetic properties the method appears promising, simple, and entirely inocuous to the cells.

  5. Phase transfer of 1- and 2-dimensional Cd-based nanocrystals

    NASA Astrophysics Data System (ADS)

    Kodanek, Torben; Banbela, Hadeel M.; Naskar, Suraj; Adel, Patrick; Bigall, Nadja C.; Dorfs, Dirk

    2015-11-01

    In this work, luminescent CdSe@CdS dot-in-rod nanocrystals, CdSe@CdS/ZnS nanorods as well as CdSe-CdS core-crown nanoplatelets were transferred into aqueous phase via ligand exchange reactions. For this purpose, bifunctional thiol-based ligands were employed, namely mercaptoacetic acid (MAA), 3-mercaptopropionic acid (MPA), 11-mercaptoundecanoic acid (MUA) as well as 2-(dimethylamino)ethanthiol (DMAET). Systematic investigations by means of photoluminescence quantum yield measurements as well as photoluminescence decay measurements have shown that the luminescence properties of the transferred nanostructures are affected by hole traps (induced by the thiol ligands themselves) as well as by spatial insulation and passivation against the environment. The influence of the tips of the nanorods on the luminescence is, however, insignificant. Accordingly, different ligands yield optimum results for different nanoparticle samples, mainly depending on the inorganic passivation of the respective samples. In case of CdSe@CdS nanorods, the highest emission intensities have been obtained by using short-chain ligands for the transfer preserving more than 50% of the pristine quantum yield of the hydrophobic nanorods. As opposed to this, the best possible quantum efficiency for the CdSe@CdS/ZnS nanorods has been achieved via MUA. The gained knowledge could be applied to transfer for the first time 2-dimensional CdSe-CdS core-crown nanoplatelets into water while preserving significant photoluminescence (up to 12% quantum efficiency).In this work, luminescent CdSe@CdS dot-in-rod nanocrystals, CdSe@CdS/ZnS nanorods as well as CdSe-CdS core-crown nanoplatelets were transferred into aqueous phase via ligand exchange reactions. For this purpose, bifunctional thiol-based ligands were employed, namely mercaptoacetic acid (MAA), 3-mercaptopropionic acid (MPA), 11-mercaptoundecanoic acid (MUA) as well as 2-(dimethylamino)ethanthiol (DMAET). Systematic investigations by means of

  6. Effective Hydraulic Conductivity Scaling in a 2-Dimensional Geometrical Multifractal Model for Aquifer Heterogeneity

    NASA Astrophysics Data System (ADS)

    Gentry, R. W.; Perfect, E.; Sukop, M. C.

    2005-12-01

    Recent analyses of field data suggest that the spatial variation of hydraulic conductivity, K, within an aquifer may be multifractal. We investigated the implications of this finding for the scaling of effective hydraulic conductivity, , by performing numerical simulations of flow in 2-dimensional geometrical multifractal K fields. A theoretical framework for generating such fields is presented based on the parameters of the truncated binomial distribution, TBD. This leads to an approximate analytical expression showing that increases with increasing length scale as a power law, whose exponent, α, is determined by the TBD parameters. Five geometrical multifractal K fields were generated with different minimum length scales. Each domain was discretized using a block center grid consisting of 59,049 uniformly-spaced nodes. A unit cube aquifer was used for the numerical simulations. The boundary conditions were implemented with constant head (unit gradient) parallel planes, and corresponding zero flux planes on the normal axes. A finite difference simulation model based on MODFLOW 2000 was used, and "zone budget" was employed to calculate the flow balance. The discharge into and out of the unit cube was then used to calculate based on Darcy's law. The numerical simulations produced similar increases in with increasing length scale to those predicted by the analytical model. Nonlinear regression analyses yielded estimates of α from the numerical simulations that were within 10% of the analytical value for these fields. These simulations provide a theoretical explanation for effective hydraulic conductivity scaling in terms of multifractals. The advantage of such an approach is that the α-parameter, which controls the degree of scaling, is physically-based and can potentially be estimated from independent measurements.

  7. 2-Dimensional Strain Analysis of Regional Change in Right Ventricular Function after Treadmill Exercise

    PubMed Central

    Yoon, Se-Jung; Park, Sujung; Chung, Wook-Jin

    2016-01-01

    Background Function of right ventricle (RV) influences on symptoms and prognosis in various diseases. However the regional RV function analyzed with 2-dimensional (2D) strain echocardiography before and just after treadmill test has not been evaluated. The aim of this study was to show the change of regional RV function just after treadmill exercise with strain analysis. Methods A total of thirty eight patients who visited hospital for hypertension, chest pain or dyspnea between January 2007 and December 2010 were retrospectively analyzed (men, 47.4%; mean age, 54.9 ± 7.2 years). Treadmill exercise test and pre and post echocardiography were performed. 2D strain echocardiography was analyzed off line in RV free wall and septum. Results Mean exercise duration was 737 ± 132 sec. Tissue velocity in lateral tricuspid annulus is significantly increased in post exercise (initial, 10.5 ± 2.4 cm/sec vs. post exercise, 12.2 ± 1.8 cm/sec, p = 0.006). Systolic strain of RV free wall apex and mid portion were significantly changed in post exercise stage (free wall apex, -18.2 ± 7.6% vs. -22.3 ± 5.8%, p = 0.010; free wall mid, -14.1 ± 6.7% vs. -22.6 ± 6.8%, p = 0.022). Conclusion 2D strain imaging provides a precise tool to quantify regional RV function and reveals a characteristic regional pattern of RV after treadmill exercise. PMID:27081442

  8. Further characterization of filarial antigens by SDS polyacrylamide gel electrophoresis

    PubMed Central

    Dissanayake, S.; Galahitiyawa, S. C.; Ismail, M. M.

    1983-01-01

    SDS (sodium dodecyl sulfate)-polyacrylamide gel electrophoresis of an antigen isolated from sera of Wuchereria bancrofti-infected patients and Setaria digitata antigen SD2-4 is reported. Both antigens showed carbohydrate (glycoprotein) staining. The W. bancrofti antigen had an apparent relative molecular mass of 35 000 while the S. digitata antigen SD2-4 migrated at the marker dye position on SDS-polyacrylamide gel electrophoresis. SDS treatment of these antigens did not abolish the precipitation reaction with antibody. In the case of W. bancrofti antigen, SDS treatment probably exposed hitherto hidden antigen epitopes. PMID:6354508

  9. Separation of DNA by free flow electrophoresis in space.

    PubMed

    Kobayashi, H; Ishii, N

    2001-10-01

    Free flow electrophoresis of a nematode C. elegans DNA was carried out on the space shuttle flight in International Microgravity Laboratory No. 2 (IML-2)., 1994. We selected the C. elegans DNA as the sample of the space experiment for free flow electrophoresis separation. This worm is a useful animal for the study of development and behavior by genetic analysis, and is a good candidate for a complete DNA sequence analysis because the haploid genome size consists of approximately 100 Mb (megabase) distributed on the six chromosomes, only 1/30 of human genome size. (Recently, the entire genomic DNA (97 Mb) sequences of C. elegans have been completed at the last December, 1998, as the first organism of multicellular system. In addition, many of the genes in C. elegans have extensive similarity to their mammalian counterparts. It may be possible to use the technology of free flow electrophoresis to contribute to the DNA analysis of the eukaryote. In this mission we attempted to make real-time communication between the space shuttle and on the ground during the electrophoresis processing. Because the separation tendency of the sample in space was not predicted so perfectly, the requests to the crews of the space shuttle was needed for selecting the fractions to be separated. According to the three dimensional electropherogram (3DEP) figured out the electrophoresis behavior, we were able to recover those fractions and kept in the deep freezer until landing. This real-time monitoring of the electrophoresis was the first evidence during space electrophoresis experiments which was started at Apollo 14, 1974. Unfortunately, some troubles occurred by contamination of bubbles in space nevertheless the post flight analyses of the fractionated samples were succeeded. The DNAs estimated by the DNA probes were not isolated one but the migration tendencies were differ from each other. It was clear that certain parts of the process in this study are advanced; that is, the precise

  10. Construction and evaluation of a capillary electrophoresis DNA sequencer

    SciTech Connect

    Drossman, H.

    1992-01-01

    This dissertation describes the construction and evaluation of an automated DNA sequencer using capillary gel electrophoresis (CGE) for separating single-strand DNA fragments and a fluorescence detector for analyzing labeled fragments. Theories governing the electrophoretic separation of DNA, dispersion processes in CGE and high sensitivity fluorescence detection are reviewed. The CGE DNA sequencer is compared with current DNA sequencing instruments and with projections of future DNA sequencing instruments. Parameters affecting the limits of detection, DNA sample loading, sample mobility and resolution are evaluated. Predictions for the future of capillary electrophoresis for large-scale sequencing projects are presented.

  11. EOS production on the Space Station. [Electrophoresis Operations/Space

    NASA Technical Reports Server (NTRS)

    Runge, F. C.; Gleason, M.

    1986-01-01

    The paper discusses a conceptual integration of the equipment for EOS (Electrophoresis Operations/Space) on the Space Station in the early 1990s. Electrophoresis is a fluid-constituent separation technique which uses forces created by an electrical field. Aspects covered include EOS equipment and operations, and Space Station installations involving a pressurized module, a resupply module, utility provisions and umbilicals and crew involvement. Accommodation feasibility is generally established, and interfaces are defined. Space Station production of EOS-derived pharmaceuticals will constitute a significant increase in capability compared to precursor flights on the Shuttle in the 1980s.

  12. Definition of performance specifications for automated Analytical Electrophoresis Facility (AAEF)

    NASA Technical Reports Server (NTRS)

    Brooks, D. E.

    1976-01-01

    In order to provide specifications for the automated Analytical Electrophoresis Facility (AAEF) that would satisfy the broadest variety of demands of a future user community, a survey was carried out of all those people who were identified as having published papers on cell electrophoresis in the past four years. A computer search was conducted of the relevant literature from which a list of 87 investigators was derived and defined as the user community for purposes of the mailing. A questionnaire was developed covering the areas of performance which required definition which was subsequently circulated to the user community. Based on the response to this survey performance specifications were assembled.

  13. Automatic multiple-sample applicator and electrophoresis apparatus

    NASA Technical Reports Server (NTRS)

    Grunbaum, B. W. (Inventor)

    1977-01-01

    An apparatus for performing electrophoresis and a multiple-sample applicator is described. Electrophoresis is a physical process in which electrically charged molecules and colloidal particles, upon the application of a dc current, migrate along a gel or a membrane that is wetted with an electrolyte. A multiple-sample applicator is provided which coacts with a novel tank cover to permit an operator either to depress a single button, thus causing multiple samples to be deposited on the gel or on the membrane simultaneously, or to depress one or more sample applicators separately by means of a separate button for each applicator.

  14. Gel Electrophoresis of Gold-DNA Nano-Conjugates

    SciTech Connect

    Pellegrino, T.; Sperling, R.A.; Alivisatos, A.P.; Parak, W.J.

    2006-01-10

    Single stranded DNA of different lengths and different amounts was attached to colloidal phosphine stabilized Au nanoparticles. The resulting conjugates were investigated in detail by a gel electrophoresis study based on 1200 gels. We demonstrate how these experiments help to understand the binding of DNA to Au particles. In particular we compare specific attachment of DNA via gold-thiol bonds with nonspecific adsorption of DNA. The maximum number of DNA molecules that can be bound per particle was determined. We also compare several methods to used gel electrophoresis for investigating the effective diameter of DNA-Au conjugates, such as using a calibration curve of particles with known diameters and Ferguson plots.

  15. Automatic registration and segmentation algorithm for multiple electrophoresis images

    NASA Astrophysics Data System (ADS)

    Baker, Matthew S.; Busse, Harald; Vogt, Martin

    2000-06-01

    We present an algorithm for registering, segmenting and quantifying multiple scanned electrophoresis images. (2D gel) Electrophoresis is a technique for separating proteins or other macromolecules in organic material according to net charge and molecular mass and results in scanned grayscale images with dark spots against a light background marking the presence of such macromolecules. The algorithm begins by registering each of the images using a non-rigid registration algorithm. The registered images are then jointly segmented using a Markov random field approach to obtain a single segmentation. By using multiple images, the effect of noise is greatly reduced. We demonstrate the algorithm on several sets of real data.

  16. GELBANK : A database of annotated two-dimensional gel electrophoresis patterns of biological systems with completed genomes.

    SciTech Connect

    Babnigg, G.; Giometti, C. S.; Biosciences Division

    2004-01-01

    GELBANK is a publicly available database of two-dimensional gel electrophoresis (2DE) gel patterns of proteomes from organisms with known genome information (available at and ftp://bioinformatics.anl.gov/gelbank/). Currently it includes 131 completed, mostly microbial proteomes available from the National Center for Biotechnology Information. A web interface allows the upload of 2D gel patterns and their annotation for registered users. The images are organized by species, tissue type, separation method, sample type and staining method. The database can be queried based on protein or 2DE-pattern attributes. A web interface allows registered users to assign molecular weight and pH gradient profiles to their own 2D gel patterns as well as to link protein identifications to a given spot on the pattern. The website presents all of the submitted 2D gel patterns where the end-user can dynamically display the images or parts of images along with molecular weight, pH profile information and linked protein identification. A collection of images can be selected for the creation of animations from which the user can select sub-regions of interest and unlimited 2D gel patterns for visualization. The website currently presents 233 identifications for 81 gel patterns for Homo sapiens, Methanococcus jannaschii, Pyro coccus furiosus, Shewanella oneidensis, Escherichia coli and Deinococcus radiodurans.

  17. Capillary electrophoresis for regulatory analysis: is it ready? Problems encountered during an interlaboratory study utilizing capillary electrophoresis.

    PubMed

    Mopper, B; Sciacchitano, C J

    1997-01-01

    Recently, the Northeast Regional Laboratory of the Food and Drug Administration conducted a collaborative study on the method to determine histamine in tuna by capillary electrophoresis. This study, under the guidance of the AOAC International, was the first of its kind involving the use of CE and was undertaken to establish official monograph status. The results of the collaborative study, however, were highly variable. Percent recovery data obtained by 11 collaborators ranged from 54 to 281%, with an average of 111%. In this paper, problems with reproducibility, and factors such as low sensitivity, noisy baselines, and variable migration times encountered by collaborators using various commercial capillary electrophoresis units are discussed. PMID:9624572

  18. Proteomic study of a model causative agent of harmful red tide, Prorocentrum triestinum I: Optimization of sample preparation methodologies for analyzing with two-dimensional electrophoresis.

    PubMed

    Chan, Leo Lai; Lo, Samuel Chun-Lap; Hodgkiss, Ivor John

    2002-09-01

    A comprehensive study to find the optimal sample preparation conditions for two-dimensional electrophoresis (2-DE) analysis of Prorocentrum triestinum, a model causative agent of harmful algal blooms (HABs) was carried out. The four major sample preparation steps for 2-DE: (a) cell disruption: i.e. sonication and homogenization with glass beads; (b) protein extraction : i.e. sequential and independent extraction procedures; (c) pre-electrophoretic treatment: these included (i) treatment with RNAase/DNAase or benzonase; (ii) ultracentrifugation to sediment large macromolecules such as DNA; (iii) desalting and concentration by ultrafiltration through a Microcon centrifugal filter device (MWCO: 3000 daltons); and (iv) desalting by a micro BioSpin chromatography column (MWCO: 6000 daltons); and (d) rehydration buffers, reducing agents and sample application in the first dimension isoelectric focussing were studied. Our results showed that sonication is easy to perform and resulted in a higher protein yield. Among the four extraction buffers, the urea containing buffers resulted in the extraction of the highest amount of protein while tris(hydroxymethyl)aminomethane buffers and trichloroacetic acid (TCA)/acetone precipitation allowed detection of a higher number of protein species (i.e. protein spots). Desalting by BioSpin and ultrafiltration have improved the 2-DE resolution of the water soluble fraction but have less effect on urea containing fractions. TCA/acetone precipitation was able to desalt all protein fractions independent of the extraction media, however extended exposure to this low pH medium has caused protein modification. Introduction of either DNase/RNase or benzonase treatment did not improve the discriminatory power of the 2-DE but this treatment did yield 2-DE with the clearest background. Proteolytic digestion was inhibited by addition of a protease inhibitor cocktail. Taken overall, a combination of sequential extraction and desalting by Bio

  19. Application of capillary electrophoresis in agricultural and soil chemistry research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As a modern analytical technique, capillary electrophoresis (CE) has become an attractive method for characterizing molecules wit high structural complexity and a wide range of molecular weights. CE can be used to analyze many natural chemical components such as acids, biogenic amines, peptides, pro...

  20. Gel Electrophoresis--The Easy Way for Students

    ERIC Educational Resources Information Center

    VanRooy, Wilhelmina; Sultana, Khalida

    2010-01-01

    This article describes a simple, inexpensive, easy to conduct gel-electrophoresis activity using food dyes. It is an alternative to the more expensive counterparts which require agarose gel, DNA samples, purchased chamber and Tris-borate-EDTA buffer. We suggest some learning activities for senior biology students along with comments on several…

  1. An Inexpensive Device for Capillary Electrophoresis with Fluorescence Detection

    ERIC Educational Resources Information Center

    Anderson, Greg; Thompson, Jonathan E.; Shurrush, Khriesto

    2006-01-01

    We describe an inexpensive device for performing capillary electrophoresis (CE) separations with fluorescence detection. As a demonstration of the device's utility we have determined the mass of riboflavin in a commercially available dietary supplement. The device allows for separation of riboflavin in [asymptotically equivalent to] 100 s with a…

  2. Capillary electrophoresis application in metal speciation and complexation characterization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Capillary electrophoresis is amenable to the separation of metal ionic species and the characterization of metal-ligand interactions. This book chapter reviews and discusses three representative case studies in applications of CE technology in speciation and reactions of metal with organic molecules...

  3. Pulsed-field gel electrophoresis typing of Staphylococcus aureus isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pulsed-field gel electrophoresis (PFGE) is the most applied and effective genetic typing method for epidemiological studies and investigation of foodborne outbreaks caused by different pathogens, including Staphylococcus aureus. The technique relies on analysis of large DNA fragments generated by th...

  4. Continuous-flow electrophoresis: Membrane-associated deviations of buffer pH and conductivity

    NASA Technical Reports Server (NTRS)

    Smolka, A. J. K.; Mcguire, J. K.

    1978-01-01

    The deviations in buffer pH and conductivity which occur near the electrode membranes in continuous-flow electrophoresis were studied in the Beckman charged particle electrophoresis system and the Hanning FF-5 preparative electrophoresis instrument. The nature of the membranes separating the electrode compartments from the electrophoresis chamber, the electric field strength, and the flow rate of electrophoresis buffer were all found to influence the formation of the pH and conductivity gradients. Variations in electrode buffer flow rate and the time of electrophoresis were less important. The results obtained supported the hypothesis that a combination of Donnan membrane effects and the differing ionic mobilities in the electrophoresis buffer was responsible for the formation of the gradients. The significance of the results for the design and stable operation of continuous-flow electrophoresis apparatus was discussed.

  5. An investigation of cutting mechanics in 2 dimensional ultrasonic vibration assisted milling toward chip thickness and chip formation

    NASA Astrophysics Data System (ADS)

    Rasidi, I. I.; Rafai, N. H.; Rahim, E. A.; Kamaruddin, S. A.; Ding, H.; Cheng, K.

    2015-12-01

    The purpose of this paper is to investigate the effects of 2 dimensional Ultrasonic Vibration Assisted Milling (UVAM) cutting mechanics, considering tool path trajectory and the effect on the chip thickness. The theoretical modelling of cutting mechanics is focused by considering the trajectory of the tool locus into the workpiece during the machining. The studies found the major advantages of VAM are come from the intermittent tool tip interaction phenomena between cutting tool and workpiece. The reduction of thinning chip thickness formations can be identifying advantages from vibration assisted milling in 2 dimensional. The finding will be discussing the comparison between conventional machining the potential of the advantages toward the chip thickness and chip formation in conclusion.

  6. Advancements to the theory of free solution electrophoresis of polyelectrolytes

    NASA Astrophysics Data System (ADS)

    McCormick, Laurette

    Capillary electrophoresis (CE) is the workhorse of countless analytical laboratories and is used routinely in various industries including pharmaceutical, forensic and clinical applications. Basically, CE is a method for separating charged molecular species in a buffer-filled capillary by the application of an electric field; the analytes move from one end of the capillary to the detector at the other end at speeds determined by their charge, size and shape. Generally, in free solution CE uniformly charged polyelectrolytes (such as DNA) are free-draining, meaning that their speed is independent of their size. Hence, until recently, a gel or other sieving medium has been necessary for the separation of polyelectrolytes; however, modifying uniformly charged polymers on the molecular level, via conjugation to uncharged polymers, allows for separation in free solution CE. In this thesis, advancements to the theory of free solution electrophoresis of polyelectrolytes, in particular, to the theories for two new free solution electrophoresis methods relying on conjugation, are presented. The first method, called End Labelled Free Solution Electrophoresis (ELFSE), can be used to sequence DNA, a negatively charged polymer in solution. Two different means of improving the resolution of ELFSE are predicted, one based on the molecular end effect, the other based on using a controlled electro-osmotic flow. In addition, a theory for the segregation of the DNA and label coils in ELFSE is presented. The second method is called Free Solution Conjugate Electrophoresis (FSCE); it allows for characterization of a sample of neutral polymers differing in length. The relevant theory, developed herein, elucidates how to accurately determine the molar mass distribution of the sample through FSCE measurements. In addition, supporting theories are developed that clarify the correct equation for the diffusion coefficient of molecules undergoing free solution electrophoresis, as well as

  7. An Optimized Trichloroacetic Acid/Acetone Precipitation Method for Two-Dimensional Gel Electrophoresis Analysis of Qinchuan Cattle Longissimus Dorsi Muscle Containing High Proportion of Marbling

    PubMed Central

    Hao, Ruijie; Adoligbe, Camus; Jiang, Bijie; Zhao, Xianlin; Gui, Linsheng; Qu, Kaixing; Wu, Sen; Zan, Linsen

    2015-01-01

    Longissimus dorsi muscle (LD) proteomics provides a novel opportunity to reveal the molecular mechanism behind intramuscular fat deposition. Unfortunately, the vast amounts of lipids and nucleic acids in this tissue hampered LD proteomics analysis. Trichloroacetic acid (TCA)/acetone precipitation is a widely used method to remove contaminants from protein samples. However, the high speed centrifugation employed in this method produces hard precipitates, which restrict contaminant elimination and protein re-dissolution. To address the problem, the centrifugation precipitates were first grinded with a glass tissue grinder and then washed with 90% acetone (TCA/acetone-G-W) in the present study. According to our result, the treatment for solid precipitate facilitated non-protein contaminant removal and protein re-dissolution, ultimately improving two-dimensional gel electrophoresis (2-DE) analysis. Additionally, we also evaluated the effect of sample drying on 2-DE profile as well as protein yield. It was found that 30 min air-drying did not result in significant protein loss, but reduced horizontal streaking and smearing on 2-DE gel compared to 10 min. In summary, we developed an optimized TCA/acetone precipitation method for protein extraction of LD, in which the modifications improved the effectiveness of TCA/acetone method. PMID:25893432

  8. Rapid DNA sequencing by horizontal ultrathin gel electrophoresis.

    PubMed Central

    Brumley, R L; Smith, L M

    1991-01-01

    A horizontal polyacrylamide gel electrophoresis apparatus has been developed that decreases the time required to separate the DNA fragments produced in enzymatic sequencing reactions. The configuration of this apparatus and the use of circulating coolant directly under the glass plates result in heat exchange that is approximately nine times more efficient than passive thermal transfer methods commonly used. Bubble-free gels as thin as 25 microns can be routinely cast on this device. The application to these ultrathin gels of electric fields up to 250 volts/cm permits the rapid separation of multiple DNA sequencing reactions in parallel. When used in conjunction with 32P-based autoradiography, the DNA bands appear substantially sharper than those obtained in conventional electrophoresis. This increased sharpness permits shorter autoradiographic exposure times and longer sequence reads. Images PMID:1870968

  9. Transverse agarose pore gradient gel electrophoresis of DNA.

    PubMed

    Fawcett, J S; Wheeler, D; Chrambach, A

    1992-06-01

    Transverse agarose pore gradient gels were prepared on GelBond in the concentration range of nominally 0.2-1.5% SeaKem GTG agarose, using density stabilization by glycerol and incorporation of a dye to define the gel concentration at each point on the pore gradient gel. The distribution of the dye was evaluated by photography, video-acquisition and digitization of the gradient mixture and by densitometry of the gel. The gel was applied to the electrophoresis of a 1-kb standard ladder of DNA fragments, using standard submarine apparatus. The method extends to agarose gel electrophoresis the benefits of semi-automated analysis of 'Ferguson curves' described in application to polyacrylamide gel by Wheeler et al. (J. Biochem. Biophys. Methods 24, 171-180). PMID:1640052

  10. Separation of DNA restriction fragments using capillary electrophoresis

    SciTech Connect

    Chan, K.C.; Whang, Chenwen; Yeung, E.S. )

    1993-01-01

    Gel-filled and non-gel' capillary electrophoresis (CE) have been applied to the separation of various DNA restriction fragments. 30% HydroLink gel, polymerized inside a 75[mu]m i.d. fused-silica capillary, was used in the gel-filled CE. Primary results show that the HL capillary gel was simple to cast, and its stability was reasonably good under the running conditions. In the non-gel CE experiment, a buffer containing the sieving additive hydroxypropylmethyl cellulose was used to affect the size-dependent separation. The use of GC capillaries eliminates the inconvenience of separately coating the capillary walls for efficient non-gel separation. Finally, the authors demonstrate that it is feasible to detect native DNA fragments using indirect fluorometry in non-gel capillary electrophoresis.

  11. Development of coatings to control electroosmosis in zero gravity electrophoresis

    NASA Technical Reports Server (NTRS)

    Krupnick, A. C.

    1974-01-01

    A major problem confronting the operation of free fluid electrophoresis in zero gravity is the control of electrokinetic phenomena and, in particular, electroosmosis. Due to the severity of counter flow as a result of electroosmosis, the electrical potential developed at the surface of shear must be maintained at near, or as close, to zero millivolts as possible. Based upon this investigation, it has been found that the amount of bound water or the degree of hydroxylation plays a major role in the control of this phenomenon. Based upon tests employing microcapillary electrophoresis, it has been found that gamma amino propyl trihydroxysilane produced a coating which provides the lowest potential (about 3.86 mV) at the surface of shear between the stationary and mobile layers.

  12. Purification of DNA Oligos by denaturing polyacrylamide gel electrophoresis (PAGE).

    PubMed

    Lopez-Gomollon, Sara; Nicolas, Francisco Esteban

    2013-01-01

    After chemical synthesis, the oligonucleotide preparation contains the desired full-length oligonucleotide but also all of the DNA molecules that were aborted during each cycle in the synthesis, and the by-products generated during the chemical reactions. The purification of oligonucleotides is a critical step for demanding applications where the exact length or sequence of the oligonucleotide is important, or for oligonucleotides longer than 50 bases. There are several methods of increasing oligonucleotide purity, the choice of which will depend on modifications of the oligonucleotides and their intended use. Polyacrylamide gel purification (PAGE purification) is the method of choice when the highest percentage of full-length oligonucleotide is desired. This chapter describes a protocol for oligonucleotide purification using denaturing polyacrylamide gel electrophoresis, and includes oligonucleotide preparation, polyacrylamide gel electrophoresis, and purification from the gel slice by two different methods: by diffusion or by electroelution. This chapter also includes recommendations as well as protocol advice. PMID:24011037

  13. Effects of coating rectangular microscopic electrophoresis chamber with methylcellulose

    NASA Technical Reports Server (NTRS)

    Plank, L. D.

    1985-01-01

    One of the biggest problems in obtaining high accuracy in microscopic electrophoresis is the parabolic flow of liquid in the chamber due to electroosmotic backflow during application of the electric field. In chambers with glass walls the source of polarization leading to electroosmosis is the negative charge of the silicare and other ions that form the wall structure. It was found by Hjerten, who used a rotating 3.0 mm capillary tube for free zone electrophoresis, that precisely neutralizing this charge was extremely difficult, but if a neutral polymer matrix (formaldehyde fixed methylcellulose) was formed over the glass (quartz) wall the double layer was displaced and the viscosity at the shear plane increased so that electroosmotic flow could be eliminated. Experiments were designed to determine the reliability with which methylcellulose coating of the Zeiss Cytopherometer chamber reduced electroosmotic backflow and the effect of coating on the accuracy of cell electrophoretic mobility (EPN) determinations. Fixed rat erythrocytes (RBC) were used as test particles.

  14. Imaging Catalytic Surfaces by Multiplexed Capillary Electrophoresis With Absorption Detection

    SciTech Connect

    Michael Christodoulou

    2002-08-27

    A new technique for in situ imaging and screening heterogeneous catalysts by using multiplexed capillary electrophoresis with absorption detection was developed. By bundling the inlets of a large number of capillaries, an imaging probe can be created that can be used to sample products formed directly from a catalytic surface with high spatial resolution. In this work, they used surfaces made of platinum, iron or gold wires as model catalytic surfaces for imaging. Various shapes were recorded including squares and triangles. Model catalytic surfaces consisting of both iron and platinum wires in the shape of a cross were also imaged successfully. Each of the two wires produced a different electrochemical product that was separated by capillary electrophoresis. Based on the collected data they were able to distinguish the products from each wire in the reconstructed image.

  15. Detection of Hb Constant Spring by a capillary electrophoresis method.

    PubMed

    Liao, Can; Zhou, Jian-Ying; Xie, Xing-Mei; Li, Jian; Li, Ru; Li, Dong-Zhi

    2010-01-01

    Hb Constant Spring [Hb CS; alpha142, Term-->Gln (TAA>CAA in alpha2)] is the most prevalent nondeletional alpha-thalassemia (alpha-thal) anomaly in southern China. In conjunction with alpha(0)-thal, it can cause severe Hb H (beta(4)) disease. The present study was done to evaluate the efficiency of two diagnostic methods in detecting Hb CS. Automated high performance liquid chromatography (HPLC) and Sebia Capillarys 2, a capillary electrophoresis method, were applied to blood samples from 21 individuals with Hb CS trait. Of the 21 cases, all (100%) were detected by capillary electrophoresis, whereas only 16 (76.2%) were detected by HPLC. We concluded that the Sebia Capillarys 2 is the preferred method for Hb CS screening. PMID:20353355

  16. Some aspects of continuous flow electrophoresis in microgravity

    NASA Astrophysics Data System (ADS)

    Biscans, B.; Hennequin, J. C.; Bertrand, J.

    Zone electrophoresis is a highly efficient method of separating biological products. It is based on the differences of mobilities of ionized particles in an electric field. During the separation complications arise due to electro-osmosis or thermal convection generated by Joule heating. This paper analyses the hydrodynamical running of a continuous flow zone electrophoresis cell and shows the influence of specific parameters on the separation. The modelling of the flow structure of the buffer solution is developed. The equations and the corresponding boundary conditions are solved by finite difference method. The model established provides knowledge of the hydrodynamical perturbations generated by electro-osmosis and thermal free convection. The case of microgravity is considered.

  17. Acupuncture Sample Injection for Microchip Capillary Electrophoresis and Electrokinetic Chromatography.

    PubMed

    Ha, Ji Won; Hahn, Jong Hoon

    2016-05-01

    A simple nanoliter-scale injection technique was developed for polydimethylsiloxane (PDMS) microfluidic devices to form the well-defined sample plugs in microfluidic channels. Sample injection was achieved by performing acupuncture on a channel with a needle and applying external pressure to a syringe. This technique allowed us to achieve reproducible injection of a 3-nL segment into a microchannel for PDMS microchip-based capillary electrophoresis (CE). Capillary zone electrophoresis (CZE) and capillary electrochromatography (CEC) with bead packing were successfully performed by applying a single potential in the most simplified straight channel. The advantages of this acupuncture injection over the electrokinetic injection in microchip CE include capability of minimizing sample loss and voltage control hardware, capability of serial injections of different sample solutions into a same microchannel, capability of injecting sample plugs into any position of a microchannel, independence on sample solutions during the loading step, and ease in making microchips due to the straight channel, etc. PMID:27056036

  18. Microchip Electrophoresis at Elevated Temperatures and High Separation Field Strengths

    PubMed Central

    Mitra, Indranil; Marczak, Steven P.; Jacobson, Stephen C.

    2014-01-01

    We report free-solution microchip electrophoresis performed at elevated temperatures and high separation field strengths. We used microfluidic devices with 11-cm long separation channels to conduct separations at temperatures between 22 (ambient) and 45 °C and field strengths from 100 to 1000 V/cm. To evaluate separation performance, N-glycans were used as a model system and labeled with 8-aminopyrene-1,3,6-trisulfonic acid to impart charge for electrophoresis and render them fluorescent. Typically, increased diffusivity at higher temperatures leads to increased axial dispersion and poor separation performance; however, we demonstrate that sufficiently high separation field strengths can be used to offset the impact of increased diffusivity in order to maintain separation efficiency. Efficiencies for these free-solution separations are the same at temperatures of 25, 35, and 45 °C with separation field strengths ≥500 V/cm. PMID:24114979

  19. A simple gel electrophoresis method for separating polyhedral gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Kim, Suhee; Lee, Hye Jin

    2015-07-01

    In this paper, a simple approach to separate differently shaped and sized polyhedral gold nanoparticles (NPs) within colloidal solutions via gel electrophoresis is described. Gel running parameters for separating efficiently gold NPs including gel composition, added surfactant types and applied voltage were investigated. The plasmonic properties and physical structure of the separated NPs extracted from the gel matrix were then investigated using transmission electron microscopy (TEM) and UV-vis spectrophotometry respectively. Data analysis revealed that gel electrophoresis conditions of a 1.5 % agarose gel with 0.1 % sodium dodecyl sulfate (SDS) surfactant under an applied voltage of 100 V resulted in the selective isolation of ~ 50 nm polyhedral shaped gold nanoparticles. Further efforts are underway to apply the method to purify biomolecule-conjugated polyhedral Au NPs that can be readily used for NP-enhanced biosensing platforms.

  20. Microchip Capillary Electrophoresis with Electrochemical Detection for Monitoring Environmental Pollutants

    SciTech Connect

    Chen, Gang; Lin, Yuehe; Wang, Joseph

    2006-01-15

    This invited paper reviews recent advances and the key strategies in microchip capillary electrophoresis (CE) with electrochemical detection (ECD) for separating and detecting a variety of environmental pollutants. The subjects covered include the fabrication of microfluidic chips, sample pretreatments, ECD, typical applications of microchip CE with ECD in environmental analysis, and future prospects. It is expected that microchip CE-ECD will become a powerful tool in the environmental field and will lead to the creation of truly portable devices.

  1. The electrophoresis of transferrins in urea/polyacrylamide gels.

    PubMed Central

    Evans, R W; Williams, J

    1980-01-01

    The denaturation of transferrin by urea has been studied by (a) electrophoresis in polyacrylamide gels incorporating a urea gradient, (b) measurements of the loss of iron-binding capacity and (c) u.v. difference spectrometry. In human serum transferrin and hen ovotransferrin the N-terminal and C-terminal domains of the iron-free protein were found to denature at different urea concentrations. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 7. PMID:7213345

  2. Sheathless interface for coupling capillary electrophoresis with mass spectrometry

    SciTech Connect

    Wang, Chenchen; Tang, Keqi; Smith, Richard D.

    2014-06-17

    A sheathless interface for coupling capillary electrophoresis (CE) with mass spectrometry is disclosed. The sheathless interface includes a separation capillary for performing CE separation and an emitter capillary for electrospray ionization. A portion of the emitter capillary is porous or, alternatively, is coated to form an electrically conductive surface. A section of the emitter capillary is disposed within the separation capillary, forming a joint. A metal tube, containing a conductive liquid, encloses the joint.

  3. Optimized photonic crystal fibers supporting efficient capillary electrophoresis

    NASA Astrophysics Data System (ADS)

    Calcerrada, M.; García-Ruiz, C.; Roy, P.; Gonzalez-Herraez, M.

    2013-05-01

    In this paper we present preliminary results on the use of Photonic Crystal Fibers (PCFs) in a conventional capillary electrophoresis system to separate and detect fluorescent species. PCFs show interesting advantages over conventional capillaries for this application, including larger surface-to-volume ratio and potential for higher resolution with comparable sensitivity. Our results illustrate some of these advantages, and we point out the need for stringent tolerances in the fabrication of specific PCFs for this application.

  4. Electrophoresis of DNA on a disordered two-dimensional substrate

    NASA Astrophysics Data System (ADS)

    Olson Reichhardt, Cynthia J.; Reichhardt, Charles

    2006-03-01

    We propose a new method for electrophoretic separation of DNA in which adsorbed polymers are driven over a disordreed two-dimensional substrate which contains attractive sites for the polymers. Using simulations of a model for long polymer chains, we show that the mobility increases with polymer length, in contrast to gel electrophoresis techniques, and that separation can be achieved for a range of length scales. We demonstrate that the separation mechanism relies on excluded volume interactions between polymer segments.

  5. Human lymphocyte polymorphisms detected by quantitative two-dimensional electrophoresis

    SciTech Connect

    Goldman, D.; Merril, C.R.

    1983-09-01

    A survey of 186 soluble lymphocyte proteins for genetic polymorphism was carried out utilizing two-dimensional electrophoresis of /sup 14/C-labeled phytohemagglutinin (PHA)-stimulated human lymphocyte proteins. Nineteen of these proteins exhibited positional variation consistent with independent genetic polymorphism in a primary sample of 28 individuals. Each of these polymorphisms was characterized by quantitative gene-dosage dependence insofar as the heterozygous phenotype expressed approximately 50% of each allelic gene product as was seen in homozygotes. Patterns observed were also identical in monozygotic twins, replicate samples, and replicate gels. The three expected phenotypes (two homozygotes and a heterozygote) were observed in each of 10 of these polymorphisms while the remaining nine had one of the homozygous classes absent. The presence of the three phenotypes, the demonstration of gene-dosage dependence, and our own and previous pedigree analysis of certain of these polymorphisms supports the genetic basis of these variants. Based on this data, the frequency of polymorphic loci for man is: P . 19/186 . .102, and the average heterozygosity is .024. This estimate is approximately 1/3 to 1/2 the rate of polymorphism previously estimated for man in other studies using one-dimensional electrophoresis of isozyme loci. The newly described polymorphisms and others which should be detectable in larger protein surveys with two-dimensional electrophoresis hold promise as genetic markers of the human genome for use in gene mapping and pedigree analyses.

  6. Visualization of DNA molecules in time during electrophoresis

    NASA Technical Reports Server (NTRS)

    Lubega, Seth

    1991-01-01

    For several years individual DNA molecules have been observed and photographed during agarose gel electrophoresis. The DNA molecule is clearly the largest molecule known. Nevertheless, the largest molecule is still too small to be seen using a microscope. A technique developed by Morikawa and Yanagida has made it possible to visualize individual DNA molecules. When these long molecules are labeled with appropriate fluorescence dyes and observed under a fluorescence microscope, although it is not possible to directly visualize the local ultrastructure of the molecules, yet because they are long light emitting chains, their microscopic dynamical behavior can be observed. This visualization works in the same principle that enables one to observe a star through a telescope because it emits light against a dark background. The dynamics of individual DNA molecules migrating through agarose matrix during electrophoresis have been described by Smith et al. (1989), Schwartz and Koval (1989), and Bustamante et al. (1990). DNA molecules during agarose gel electrophoresis advance lengthwise thorough the gel in an extended configuration. They display an extension-contraction motion and tend to bunch up in their leading ends as the 'heads' find new pores through the gel. From time to time they get hooked on obstacles in the gel to form U-shaped configurations before they resume their linear configuration.

  7. Ultrasensitive plasmid mapping by high performance capillary electrophoresis.

    PubMed

    Maschke, H E; Frenz, J; Belenkii, A; Karger, B L; Hancock, W S

    1993-01-01

    This paper compares high performance capillary electrophoresis (HPCE) and conventional slab electrophoresis in mapping of four closely related plasmids with three different restriction enzymes. The plasmids express full length and truncated forms of a growth factor receptor oncogene product and were digested with HpaII, HaeIII and RsaI. The resulting oligonucleotide fragments were under 2000 base pairs in length, a size well suited to separation by HPCE with linear polyacrylamide as a sieving matrix. Plasmid mapping is an essential tool in biotechnology both for the design of an expression system and for monitoring the stability of the expression system during fermentation. HPCE can yield much higher resolution of oligonucleotides than attainable in conventional agarose gel electrophoretic procedures for plasmid mapping. In the examples described here, the HpaII digests provided the surest identification of individual plasmids in the HPCE analysis and could discriminate among all four plasmids. In conventional slab electrophoresis, however, the RsaI digests provided the best discrimination, although two of the plasmids in this system yielded essentially identical electrophoretic patterns. Hence the optimal restriction enzyme for plasmid mapping applications with HPCE may differ from that selected on the basis of conventional slab gel analysis, and the former technique can provide higher discrimination among related plasmids. The advantages of the HPCE format with respect to speed, low sample consumption and resolution are described. PMID:8354236

  8. Improved single-strand DNA sizing accuracy in capillary electrophoresis.

    PubMed Central

    Rosenblum, B B; Oaks, F; Menchen, S; Johnson, B

    1997-01-01

    Interpolation algorithms can be developed to size unknown single-stranded (ss) DNA fragments based on their electrophoretic mobilities, when they are compared with the mobilities of standard fragments of known sizes; however, sequence-specific anomalous electrophoretic migration can affect the accuracy and precision of the called sizes of the fragments. We used the anomalous migration of ssDNA fragments to optimize denaturation conditions for capillary electrophoresis. The capillary electrophoretic system uses a refillable polymer that both coats the capillary wall to suppress electro-osmotic flow and acts as the sieving matrix. The addition of 8 M urea to the polymer solution, as in slab gel electrophoresis, is insufficient to fully denature some anomalously migrating ssDNA fragments in this capillary electrophoresis system. The sizing accuracy of these fragments is significantly improved by the addition of 2-pyrrolidinone, or increased capillary temperature (60 degrees C). the effect of these two denaturing strategies is additive, and the best accuracy and precision in sizing results are obtained with a combination of chemical and thermal denaturation. PMID:9380518

  9. Microchip electrophoresis for the analysis of biological materials

    SciTech Connect

    Jacobson, S.C.; Ramsey, J.M.

    1995-12-31

    Development of instrumentation for biological and chemical analyses is geared toward extracting more information in a shorter time period at a lower cost. One avenue to achieve these ends is the miniaturization of instrumentation using microfabrication techniques. The primary advantage of such miniaturized devices is the integration of multiple functions required for a complete analysis into a single monolithic unit. Also, by using microfabrication techniques, planar devices with compact geometries enable design of parallel architectures which would handle multiple samples and necessary redundancies. Microfabricated devices that have been demonstrated primarily involve electrically driven separation techniques including capillary electrophoresis, open channel electrochromatography, and capillary gel electrophoresis. Devices that integrate chemical reactions with analysis include capillary electrophoresis with pre- and postseparation derivatization, and restriction digestions of plasmid DNA followed by size analysis of the fragments. The most notable feature of these miniaturized chemical separation devices is the speed with which analyses are performed. These devices have precise fluid control of picoliter volumes simply by proper selection of applied potentials and without the use of mechanical valves. Further motivations for miniaturization and integration of chemical analysis procedures include low void volume connections, reduction of sample and reagent volumes, and rapid diffusional mixing of reagents.

  10. RAMSES: Applied research on separation methods using space electrophoresis

    NASA Astrophysics Data System (ADS)

    Jamin Changeart, F.; Faure, F.; Sanchez, V.; Schoot, B.; Simonis, M.; Renard, A.; Collete, J. P.; Perez, D.; Val, J. M.; de Olano, A. l.

    Eight european industrial companies, the CNRS and University Paul Sabatier and CNES/ Centre National d'Etudes Spatiales collaborate on the SBS (Space Bio Separation) project which aims at demonstrating the possibility of preparing high-purity biomaterials under microgravity conditions. As a first step of SBS, the proposal of a cooperative flight of the RAMSES facility on board Spacelab during the IML-2 mission, scheduled January 1993, has been selected by NASA. RAMSES allows basic and applied research on free flow zone electrophoresis, in order to assess the influence of a low-gravity environment on the purification of biological products. Experiments will be performed by European and American scientists. The facility will be integrated in a Spacelab single rack. Using in situ diagnostics with a U.V. photometer and a cross illuminator, RAMSES investigates a wide variety of transport phenomena to better understand the basic mechanisms which govern electrophoresis method. RAMSES should be a basis for a more complete facility dedicated to the purification of biomaterials, associating various separation methods. This paper will provide an overview of this space facility RAMSES with emphasis on continuous flow zone electrophoresis technique, scientific back-ground, RAMSES experimental program, RAMSES main functions and an overall description of the RAMSES main units.

  11. Microfabricated capillary electrophoresis amino acid chirality analyzer for extraterrestrial exploration

    NASA Technical Reports Server (NTRS)

    Hutt, L. D.; Glavin, D. P.; Bada, J. L.; Mathies, R. A.

    1999-01-01

    Chiral separations of fluorescein isothiocyanate-labeled amino acids have been performed on a microfabricated capillary electrophoresis chip to explore the feasibility of using such devices to analyze for extinct or extant life signs in extraterrestrial environments. The test system consists of a folded electrophoresis channel (19.0 cm long x 150 microns wide x 20 microns deep) that was photolithographically fabricated in a 10-cm-diameter glass wafer sandwich, coupled to a laser-excited confocal fluorescence detection apparatus providing subattomole sensitivity. Using a sodium dodecyl sulfate/gamma-cyclodextrin pH 10.0 carbonate electrophoresis buffer and a separation voltage of 550 V/cm at 10 degrees C, baseline resolution was observed for Val, Ala, Glu, and Asp enantiomers and Gly in only 4 min. Enantiomeric ratios were determined for amino acids extracted from the Murchison meteorite, and these values closely matched values determined by HPLC. These results demonstrate the feasibility of using microfabricated lab-on-a-chip systems to analyze extraterrestrial samples for amino acids.

  12. Phylogenetic reconstruction of South American felids defined by protein electrophoresis.

    PubMed

    Slattery, J P; Johnson, W E; Goldman, D; O'Brien, S J

    1994-09-01

    Phylogenetic associations among six closely related South American felid species were defined by changes in protein-encoding gene loci. We analyzed proteins isolated from skin fibroblasts using two-dimensional electrophoresis and allozymes extracted from blood cells. Genotypes were determined for multiple individuals of ocelot, margay, tigrina, Geoffroy's cat, kodkod, and pampas cat at 548 loci resolved by two-dimensional electrophoresis and 44 allozyme loci. Phenograms were constructed using the methods of Fitch-Margoliash and neighbor-joining on a matrix of Nei's unbiased genetic distances for all pairs of species. Results of a relative-rate test indicate changes in two-dimensional electrophoresis data are constant among all South American felids with respect to a hyena outgroup. Allelic frequencies were transformed to discrete character states for maximum parsimony analysis. Phylogenetic reconstruction indicates a major split occurred approximately 5-6 million years ago, leading to three groups within the ocelot lineage. The earliest divergence led to Leopardus tigrina, followed by a split between an ancestor of an unresolved trichotomy of three species (Oncifelis guigna, O. geoffroyi, and Lynchailuris colocolo) and a recent common ancestor of Leopardus pardalis and L. wiedii. The results suggest that modern South American felids are monophyletic and evolved rapidly after the formation of the Panama land bridge between North and South America. PMID:7932791

  13. Hybridization thermodynamics of DNA oligonucleotides during microchip capillary electrophoresis.

    PubMed

    Wynne, Thomas M; McCallum, Christopher; Del Bonis-O'Donnell, Jackson Travis; Crisalli, Pete; Pennathur, Sumita

    2015-03-01

    Capillary electrophoresis (CE) is a powerful analytical tool for performing separations and characterizing properties of charged species. For reacting species during a CE separation, local concentrations change leading to nonequilibrium conditions. Interpreting experimental data with such nonequilibrium reactive species is nontrivial due to the large number of variables involved in the system. In this work we develop a COMSOL multiphysics-based numerical model to simulate the electrokinetic mass transport of short interacting ssDNAs in microchip capillary electrophoresis. We probe the importance of the dissociation constant, K(D), and the concentration of DNA on the resulting observed mobility of the dsDNA peak, μ(w), by using a full sweep of parametric simulations. We find that the observed mobility is strongly dependent on the DNA concentration and K(D), as well as ssDNA concentration, and develop a relation with which to understand this dependence. Furthermore, we present experimental microchip capillary electrophoresis measurements of interacting 10 base ssDNA and its complement with changes in buffer ionic strength, DNA concentration, and DNA sequence to vary the system equilibria. We then compare our results to thermodynamically calculated K(D) values. PMID:25634338

  14. Automated Lab-on-a-Chip Electrophoresis System

    NASA Technical Reports Server (NTRS)

    Willis, Peter A.; Mora, Maria; Greer, Harold F.; Fisher, Anita M.; Bryant, Sherrisse

    2012-01-01

    Capillary electrophoresis is an analytical technique that can be used to detect and quantify extremely small amounts of various biological molecules. In the search for biochemical traces of life on other planets, part of this search involves an examination of amino acids, which are the building blocks of life on Earth. The most sensitive method for detecting amino acids is the use of laser induced fluorescence. However, since amino acids do not, in general, fluoresce, they first must be reacted with a fluorescent dye label prior to analysis. After this process is completed, the liquid sample then must be transported into the electrophoresis system. If the system is to be reused multiple times, samples must be added and removed each time. In typical laboratories, this process is performed manually by skilled human operators using standard laboratory equipment. This level of human intervention is not possible if this technology is to be implemented on extraterrestrial targets. Microchip capillary electrophoresis (CE) combined with laser induced fluorescence detection (LIF) was selected as an extremely sensitive method to detect amino acids and other compounds that can be tagged with a fluorescent dye. It is highly desirable to package this technology into an integrated, autonomous, in situ instrument capable of performing CE-LIF on the surface of an extraterrestrial body. However, to be fully autonomous, the CE device must be able to perform a large number of sample preparation and analysis operations without the direct intervention of a human.

  15. [Does bilirubin interfere with capillary electrophoresis of serum proteins?].

    PubMed

    Hellara, Ilhem; Fekih, Ons; Triki, Sonia; Elmay, Ahlem; Neffati, Fadoua; Najjar, Mohamed Fadhel

    2014-01-01

    Capillary electrophoresis of serum proteins is a fast, reliable and simple technique, but many interference exist. The objective of our work is to study the interference of bilirubin on this technique; 70 icteric sera were analysed on Capillarys ™ (Sebia). A second electrophoresis was performed on 40 samples after bilirubin photodegradation. The bilirubin and serum proteins were determinated respectively by Jendrassik and Grof and biuret methods on Konélab 20i ™ (Thermo Electron Corporation). We found abnormal spreading of the albumin fraction of the anode side wich constitute sometimes an isolated fraction in the traditional area of pre-albumin migration. This fraction varies from 2.0 ± 2.0% (0.0 to 7.3%) or 0.98 ± 1.53 g/L (0 to 5.3 g/L) and it seems to be related to the direct bilirubin since, following overloading sera with a solution of bilirubin, no further fraction was recovered. An average decrease of bilirubin after photodegradation of 58 ± 17% (26-89%) is followed by a decrease in the same order 64 ± 38% (10-100%) of the additional fraction. Acetate cellulose electrophoresis of the same samples showed no variation. The high bilirubin levels seem modify slightly the electrophoretic profile. However the impact of the interference on the interpretation of electrophoretic trace is negligible. PMID:24492101

  16. ``Superfast'' and ``Hyperfast'' Electrophoresis in DC and AC Electric Fields

    NASA Astrophysics Data System (ADS)

    Demekhin, Evgeny; Korovyakovsky, Alex

    2006-11-01

    Movement of a small conducting spherical granule in an electrolyte solution under force of DC and AC fields is considered. The problem is described by strongly coupled nonlinear PDE system. The fact that it has two small parameters, the ratio of the ion double layer to the diffusion layer and the ratio of the diffusion layer to the granule's diameter, makes the problem unique and extremely difficult to solve. This is the reason why only solutions for some particular cases have been known. In this work for the first time, combining asymptotic and numerical methods, a complete theory of electrophoresis in DC and AC fields is developed. By special decomposition method the system is transformed to new variables. Analytical solution in the inner region results in the nonlinear Smoluchowski slip velocity. In the intermediate region convection-diffusion equation is solved numerically. In tern, the intermediate solution is matched with the outer solution of Laplace equation to complete the statement. For a strong DC field (``superfast'' electrophoresis) the theory predicts, in agreement with experiments, the granule's velocity to be proportional to the granule's size and squared external field; there is a large elongated vortex behind the granule and a small one near its equator. There is an excellent agreement with available experimental data. Granule's velocity for AC field becomes even larger than for DC, it has a maximum with respect to the field's frequency (``hyperfast'' electrophoresis).

  17. Electrophoresis of Ion Containing Polymers in Microfluidic Applications

    NASA Astrophysics Data System (ADS)

    Chow, Andrea

    2004-03-01

    Microfluidic technology offers the benefits of miniaturization, integration, and automation that can lead to faster analysis with higher data quality and higher sample throughput. One of the first microfluidic systems commercialized is for biomolecular sizing using the principles of gel electrophoresis. In these chips, the microchannels are filled with a polymer solution at a concentration above the entanglement threshold. For DNA and RNA sizing, the procedures of sample injection, fluorescent dye staining of the analyte, electrophoretic separation by size, and optical detection of size fractions are integrated. These microchip analyses are similar to those performed in conventional capillary electrophoresis, except that the analysis times are usually an order of magnitude shorter. For protein sizing in sodium dodecyl sulphate (SDS) micelle solutions in which the proteins are denatured, an additional step of protein destaining is also integrated onto the chip, enabling an application that has no simple analog in conventional capillary electrophoresis. The scaling laws based on polymer physics considerations dictating the sizing mechanisms and separation efficiencies for these microfluidic applications will be discussed.

  18. Capillary zone electrophoresis-mass spectrometry of peptides and proteins

    SciTech Connect

    Loo, J.A.; Udseth, H.R.; Smith, R.D.

    1989-05-01

    Capillary zone electrophoresis (CZE) is attracting extensive attention as a fast, high resolution analytical and micro-preparative separations technique for systems of biological interest. In zone electrophoresis, a column is filled with a single electrolyte having a specific conductivity. The mixture of substances to be separated is applied as a narrow band to the head of a buffer filled column in a band whose width is much less than the length of the column and at a concentration too low to affect the buffer conductivity. An electric field is then applied across the length of the column and the individual substances migrate and separate according to their net electrophoretic velocities. Zone electrophoresis carried out in small diameter (<100 ..mu..m) fused silica capillaries is a relatively new approach to the high resolution separation of aqueous samples. Very small volume samples (picoliter range) with separation efficiencies on the order of 10/sup 6/ theoretical plates for amino acids have been achieved. The method can be further enhanced by the dynamic combination of detection sensitivity and selectivity offered by mass spectrometry (MS). The on-line marriage of mass spectrometry to CZE is accomplished by an atmospheric pressure electrospray ionization source interface. Our research efforts have demonstrated that proteins with MW's greater than 100 kDa can be analyzed using a conventional quadrupole mass spectrometer with an upper m/z limit of only 1700. 6 refs.

  19. Applications of space-electrophoresis in medicine. [for cellular separations in molecular biology

    NASA Technical Reports Server (NTRS)

    Bier, M.

    1976-01-01

    The nature of electrophoresis is reviewed and potential advances realizable in the field of biology and medicine from a space electrophoresis facility are examined. The ground-based applications of electrophoresis: (1) characterization of an ionized species; (2) determination of the quantitative composition of a complex mixture; and (3) isolation of the components of a mixture, separation achieved on the basis of the difference in transport rates is reviewed. The electrophoresis of living cells is considered, touching upon the following areas: the separation of T and B lymphocytes; the genetic influence on mouse lymphocyte mobilities; the abnormal production of specific and monoclonal immunoproteins; and the study of cancer. Schematic diagrams are presented of three types of electrophoresis apparatus: the column assembly for the static electrophoresis experiment on the Apollo-Soyuz mission, the continuous flow apparatus used in the same mission and a miniaturized electrophoresis apparatus.

  20. GRAV2D: an interactive 2-1/2 dimensional gravity modeling program (user's guide and documentation for Rev. 1)

    SciTech Connect

    Nutter, C.

    1980-11-01

    GRAV2D is an interactive computer program used for modeling 2-1/2 dimensional gravity data. A forward algorithm is used to give the theoretical attraction of gravity intensity at a station due to a perturbing body given by the initial model. The resultant model can then be adjusted for a better fit by a combination of manual adjustment, one-dimensional automatic search, and Marquardt inversion. GRAV2D has an interactive data management system for data manipulation and display built around subroutines to do a forward problem, a one-dimensional direct search and an inversion. This is a user's guide and documentation for GRAV2D.

  1. Accretions of dark matter and dark energy onto (n+2)-dimensional Schwarzschild black hole and Morris-Thorne wormhole

    NASA Astrophysics Data System (ADS)

    Debnath, Ujjal

    2015-12-01

    In this work, we have studied accretion of the dark matter and dark energy onto of (n+2)-dimensional Schwarzschild black hole and Morris-Thorne wormhole. The mass and the rate of change of mass for (n+2)-dimensional Schwarzschild black hole and Morris-Thorne wormhole have been found. We have assumed some candidates of dark energy like holographic dark energy, new agegraphic dark energy, quintessence, tachyon, DBI-essence, etc. The black hole mass and the wormhole mass have been calculated in term of redshift when dark matter and above types of dark energies accrete onto them separately. We have shown that the black hole mass increases and wormhole mass decreases for holographic dark energy, new agegraphic dark energy, quintessence, tachyon accretion and the slope of increasing/decreasing of mass sensitively depends on the dimension. But for DBI-essence accretion, the black hole mass first increases and then decreases and the wormhole mass first decreases and then increases and the slope of increasing/decreasing of mass not sensitively depends on the dimension.

  2. Differentiation of acute renal failure and chronic renal failure by 2-dimensional analysis of urinary dipeptidase versus serum creatinine.

    PubMed

    Lee, S H; Kang, B Y; We, J S; Park, S K; Park, H S

    1999-03-01

    The differential diagnosis of acute renal failure (ARF) and chronic renal failure (CRF) may be possible by measuring urinary dipeptidase (Udpase) activity and serum creatinine (Scr) concentration. When the mass test of 246 individuals was examined on a 2-dimensional plot of Udpase (y-axis) versus Scr (x-axis) with the data obtained from healthy volunteers (n = 189), ARF (n = 19) and CRF (n = 38) patients, the characteristic distribution of each group was obvious. It is summarized by the mean values of healthy volunteers (1.44 +/- 0.39 mg/dL, 1.19 (0.59 mU/mL), ARF (6.04 +/- 5.04 mg/dL, 0.12 +/- 0.08 mU/mL), and CRF patients (8.72 +/- 2.93 mg/dL, 0.81 +/- 0.44 mU/mL). The healthy volunteers are distributed along the y-axis and the ARF patients the x-axis, thus separating the two groups 90 degrees apart. The CRF patients are scattered away from both x-, and y-axis. This 2-dimensional approach is thought to be very useful for the differential diagnosis of ARF suggesting Udpase as a new member of the marker enzymes of renal disease. PMID:10088177

  3. Application of multilocus enzyme gel electrophoresis to Haemophilus influenzae.

    PubMed Central

    Porras, O; Caugant, D A; Lagergård, T; Svanborg-Edén, C

    1986-01-01

    Multilocus enzyme electrophoresis was adapted to the study of Haemophilus influenzae. Protein extracts from sonicated whole bacteria were subjected to starch gel electrophoresis. After staining with substrates, the position of each isoenzyme (electromorph) was registered. Each isolate was assigned an electrophoretic type (ET) by the combination of electromorphs for the enzymes stained. Twenty-seven enzymes were tested; 12 were expressed in H. influenzae. Six enzymes were selected for subsequent study: malate dehydrogenase (MDH), phenylalanylleucine peptidase (PE2), 6-phosphogluconate dehydrogenase (6PG), adenylate kinase (AK), glucose 6-phosphate dehydrogenase (G6P), and phosphoglucose isomerase (PGI). They were polymorphic and occurred in all isolates. Six electromorphs were found for PE2, G6P, and PGI, five for MDH, four for 6PG, and three for AK. PE2, G6P, and PGI contributed most of the ET resolution (48 of 49 ETs). Multilocus enzyme electrophoresis showed several advantages over previous typing techniques. An ET could be assigned to both typable and nontypable (NT) isolates. The technique was powerful in resolving differences among isolates. The 94 isolates comprised 49 ETs, five biotypes, and six capsular types and NT isolates. Strains known to be related expressed the same ET, e.g., RAB b+ and b-, ET12; Ma a+ and a-, ET1. ET variability among type b isolates was low; 26 of 28 clinical isolates expressed ET14; 2 of 28 expressed ET13 and ET15, differing from ET14 by one electromorph each. In contrast, the 47 NT isolates comprised 38 different ETs. No ETs were shared between non-type b capsulated strains and type b or NT strains. Interestingly, five NT isolates expressed the same ET as type b strains. (iv) Strains of the same capsular type but different biotypes expressed different ETs. ET determinations will thus be useful in studying the epidemiology and evolution of H. influenzae. Images PMID:3522433

  4. Evaluation of a multicapillary electrophoresis instrument for mitochondrial DNA typing.

    PubMed

    Stewart, John E B; Aagaard, Patricia J; Pokorak, Eric G; Polanskey, Deborah; Budowle, Bruce

    2003-05-01

    Laser-induced detection of fluorescent labeled PCR products and multi-wavelength detection (i.e., multicolor analysis) enables rapid generation of mtDNA sequencing profiles. Traditionally, polyacrylamide slab gels have been used as the electrophoretic medium for mtDNA sequencing in forensic analyses. Replacement of slab gel electrophoresis with capillary electrophoresis (CE) can facilitate automation of the analytical process. Automation and high throughput can be further enhanced by using multicapillary electrophoretic systems. The use of the ABI Prism 3100 Genetic Analyzer (ABI 3100, Applied Biosystems, Foster City, CA) as well as the ABI Prism 310 Genetic Analyzer (ABI 310, Applied Biosystems, Foster City, CA) were evaluated for mtDNA sequencing capabilities and compared with sequencing results obtained on the platform currently in use in the FBI Laboratory (the ABI Prism 377 DNA Sequencer, ABI 377, Applied Biosystems, Foster City, CA). Various studies were performed to assess the utility of the ABI 3100, as well as the ABI 310 for mtDNA sequencing. The tests included: comparisons of results obtained among the ABI 3100, the ABI 310 and the ABI 377 instruments; comparisons of results obtained within and between capillary arrays; evaluation of capillary length; evaluation of sample injection time; evaluation of the resolution of mixtures/heteroplasmic samples; and evaluation of the sensitivity of detection of a minor component with reduced template on the ABI 3100. In addition, other studies were performed to improve sample preparation; these included: comparison of template suppression reagent (TSR, Applied Biosystems, Foster City, CA) versus formamide; the use of Performa DTR Gel Filtration Cartridges (Edge BioSystems Inc., Gaithersburg, MD) versus Centri-Sep Spin Columns (Princeton Separations, Adelphia, NJ) for product purification after cycle sequencing; and sample stability after denaturation. The data support that valid and reliable results can be obtained

  5. Electrophoresis and orientation of F-actin in agarose gels.

    PubMed Central

    Borejdo, J; Ortega, H

    1989-01-01

    F-Actin was electrophoresed on agarose gels. In the presence of 2 mM MgCl2 and above pH 8.5 F-actin entered 1% agarose; when the electric field was 2.1 V/cm and the pH was 8.8, F-actin migrated through a gel as a single band at a rate of 2.5 mm/h. Labeling of actin with fluorophores did not affect its rate of migration, but an increase in ionic strength slowed it down. After the electrophoresis actin was able to bind phalloidin and heavy meromyosin (HMM) and it activated Mg2+-dependent ATPase activity of HMM. The mobility of F-actin increased with the rise in pH. Acto-S-1 complex was also able to migrate in agarose at basic pH, but at a lower rate than F-actin alone. The orientation of fluorescein labeled F-actin and of fluorescein labeled S-1 which formed rigor bonds with F-actin was measured during the electrophoresis by the fluorescence detected linear dichroism method. The former showed little orientation, probably because the dye was mobile on the surface of actin, but we were able to measure the orientation of the absorption dipole of the dye bound to S-1 which was attached to F-actin, and found that it assumed an orientation largely parallel to the direction of the electric field. These results show that actin can migrate in agarose gels in the F form and that it is oriented during the electrophoresis. Images FIGURE 1 FIGURE 3 FIGURE 4 PMID:2528384

  6. Miniaturizing free-flow electrophoresis - a critical review.

    PubMed

    Kohlheyer, Dietrich; Eijkel, Jan C T; van den Berg, Albert; Schasfoort, Richard B M

    2008-03-01

    Free-flow electrophoresis (FFE) separation methods have been developed and investigated for around 50 years and have been applied not only to many types of analytes for various biomedical applications, but also for the separation of inorganic and organic substances. Its continuous sample preparation and mild separation conditions make it also interesting for online monitoring and detection applications. Since 1994 several microfluidic, miniaturized FFE devices were developed and experimentally characterized. In contrast to their large-scale counterparts microfluidic FFE (mu-FFE) devices offer new possibilities due to the very rapid separations within several seconds or below and the requirement for sample volumes in the microliter range. Eventually, these mu-FFE systems might find application in so-called lab-on-a-chip devices for real-time monitoring and separation applications. This review gives detailed information on the results so far published on mu-FFE chips, comprising its four main modes, namely free-flow zone electrophoresis (FFZE), free-flow IEF (FFIEF), free-flow ITP (FFITP), and free-flow field-step electrophoresis (FFFSE). The principles of the different FFE modes and the basic underlying theory are given and discussed with special emphasis on miniaturization. Different designs as well as fabrication methods and applied materials are discussed and evaluated. Furthermore, the separation results shown indicate that similar separation quality with respect to conventional FFE systems, as defined by the resolution and peak capacity, can be achieved with mu-FFE separations when applying much lower electrical voltages. Furthermore, innovations still occur and several approaches for hyphenated, more integrated systems have been proposed so far, some of which are discussed here. This review is intended as an introduction and early compendium for research and development within this field. PMID:18232029

  7. Recent progress in preparation and application of microfluidic chip electrophoresis

    NASA Astrophysics Data System (ADS)

    Cong, Hailin; Xu, Xiaodan; Yu, Bing; Yuan, Hua; Peng, Qiaohong; Tian, Chao

    2015-05-01

    Since its discovery in 1990, microfluidic chip electrophoresis (MCE) has allowed the development of applications with small size, fast analysis, low cost, high integration density and automatic level, which are easy to carry and have made commercialization efficient. MCE has been widely used in the areas of environmental protection, biochemistry, medicine and health, clinical testing, judicial expertise, food sanitation, pharmaceutical checking, drug testing, agrochemistry, biomedical engineering and life science. As one of the foremost fields in the research of capillary electrophoresis, MCE is the ultimate frontier to develop the miniaturized, integrated, automated all-in-one instruments needed in modern analytical chemistry. By adopting the advanced technologies of micro-machining, lasers and microelectronics, and the latest research achievements in analytical chemistry and biochemistry, the sampling, separation and detection systems of commonly used capillary electrophoresis are integrated with high densities onto glass, quartz, silicon or polymer wafers to form the MCE, which can finish the analysis of multi-step operations such as injection, enrichment, reaction, derivatization, separation, and collection of samples in a portable, efficient and super high speed manner. With reference to the different technological achievements in this area, the latest developments in MCE are reviewed in this article. The preparation mechanisms, surface modifications, and properties of different materials in MCE are compared, and the different sampling, separation and detection systems in MCE are summarized. The performance of MCE in analysis of fluorescent substance, metallic ion, sugar, medicine, nucleic acid, DNA, amino acid, polypeptide and protein is discussed, and the future direction of development is forecast.

  8. Density gradient electrophoresis of cultured human embryonic kidney cells

    NASA Technical Reports Server (NTRS)

    Plank, L. D.; Kunze, M. E.; Giranda, V.; Todd, P. W.

    1985-01-01

    Ground based confirmation of the electrophoretic heterogeneity of human embryonic kidney cell cultures, the general characterization of their electrophoretic migration, and observations on the general properties of cultures derived from electrophoretic subpopulations were studied. Cell migration in a density gradient electrophoresis column and cell electrophoretic mobility was determined. The mobility and heterogeneity of cultured human embryonic kidney cells with those of fixed rat erythrocytes as model test particle was compared. Electrophoretically separated cell subpopulations with respect to size, viability, and culture characteristics were examined.

  9. Capillary electrophoresis-electrochemical detection microchip device and supporting circuits

    DOEpatents

    Jackson, Douglas J.; Roussel, Jr., Thomas J.; Crain, Mark M.; Baldwin, Richard P.; Keynton, Robert S.; Naber, John F.; Walsh, Kevin M.; Edelen, John. G.

    2008-03-18

    The present invention is a capillary electrophoresis device, comprising a substrate; a first channel in the substrate, and having a buffer arm and a detection arm; a second channel in the substrate intersecting the first channel, and having a sample arm and a waste arm; a buffer reservoir in fluid communication with the buffer arm; a waste reservoir in fluid communication with the waste arm; a sample reservoir in fluid communication with the sample arm; and a detection reservoir in fluid communication with the detection arm. The detection arm and the buffer arm are of substantially equal length.

  10. PNEUMATIC MICROVALVE FOR ELECTROKINETIC SAMPLE PRECONCENTRATION AND CAPILLARY ELECTROPHORESIS INJECTION

    SciTech Connect

    Cong, Yongzheng; Rausch, Sarah J.; Geng, Tao; Jambovane, Sachin R.; Kelly, Ryan T.

    2014-10-27

    Here we show that a closed pneumatic microvalve on a PDMS chip can serve as a semipermeable membrane under an applied potential, enabling current to pass through while blocking the passage of charged analytes. Enrichment of both anionic and cationic species has been demonstrated, and concentration factors of ~70 have been achieved in just 8 s. Once analytes are concentrated, the valve is briefly opened and the sample is hydrodynamically injected onto an integrated microchip or capillary electrophoresis (CE) column. In contrast to existing preconcentration approaches, the membrane-based method described here enables both rapid analyte concentration as well as high resolution separations.

  11. A method for easily customizable gradient gel electrophoresis.

    PubMed

    Miller, Andrew J; Roman, Brandon; Norstrom, Eric

    2016-09-15

    Gradient polyacrylamide gel electrophoresis is a powerful tool for the resolution of polypeptides by relative mobility. Here, we present a simplified method for generating polyacrylamide gradient gels for routine analysis without the need for specialized mixing equipment. The method allows for easily customizable gradients which can be optimized for specific polypeptide resolution requirements. Moreover, the method eliminates the possibility of buffer cross contamination in mixing equipment, and the time and resources saved with this method in place of traditional gradient mixing, or the purchase of pre-cast gels, are noteworthy given the frequency with which many labs use gradient gel SDS-PAGE. PMID:27393767

  12. Capillary electrophoresis: Biotechnology for separation of DNA and chromosomes

    NASA Technical Reports Server (NTRS)

    Williams, George O., Jr.

    1994-01-01

    Electrophoresis has been used for the separation of particles, ions, and molecules for a number of years. The technology for separation and detection of the results has many applications in the life sciences. One of the major goals of the scientific community is to separate DNA molecules and intact chromosomes based upon their different lengths or number of base pairs. This may be achieved by using some of the commercially available and widely used methods, but these processes require a considerable amount of time. The challenge is to achieve separation of intact chromosomes in a short time, preferably in a matter of minutes.

  13. Capillaries for use in a multiplexed capillary electrophoresis system

    DOEpatents

    Yeung, E.S.; Chang, H.T.; Fung, E.N.

    1997-12-09

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification (``base calling``) is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations. 19 figs.

  14. High-sensitivity capillary and microchip electrophoresis using electrokinetic supercharging.

    PubMed

    Dawod, Mohamed; Chung, Doo Soo

    2011-10-01

    Electrokinetic supercharging (EKS) is considered as one of the most powerful online preconcentration techniques in electrophoresis. It combines the efficient preconcentration power of field-amplified sample injection and the exceptional selective nature of transient isotachophoresis. It has a wide range of applications to different types of analytes ranging from small ions to large proteins and DNA fragments. This comprehensive review--up to date--provides listing for all the works, developments, and advances in EKS. The review will pay particular attention to innovations, new methodologies for manipulation, challenges for improving the detection sensitivity, and various applications of EKS in capillaries and microchips. PMID:21793208

  15. Electronic imaging systems for quantitative electrophoresis of DNA

    SciTech Connect

    Sutherland, J.C.

    1989-01-01

    Gel electrophoresis is one of the most powerful and widely used methods for the separation of DNA. During the last decade, instruments have been developed that accurately quantitate in digital form the distribution of materials in a gel or on a blot prepared from a gel. In this paper, I review the various physical properties that can be used to quantitate the distribution of DNA on gels or blots and the instrumentation that has been developed to perform these tasks. The emphasis here is on DNA, but much of what is said also applies to RNA, proteins and other molecules. 36 refs.

  16. Monitoring environmental pollutants by microchip capillary electrophoresis with electrochemical detection

    SciTech Connect

    Chen, Gang; Lin, Yuehe; Wang, Joseph

    2006-01-15

    This is a review article. During the past decade, significant progress in the development of miniaturized microfluidic systems has Occurred due to the numerous advantages of microchip analysis. This review focuses on recent advances and the key strategies in microchip capillary electrophoresis (CE) with electrochemical detection (ECD) for separating and detecting a variety of environmental pollutants. The subjects covered include the fabrication of microfluidic chips, ECD, typical applications of microchip CE with ECD in environmental analysis, and future prospects. It is expected that microchip CE-ECD will become a powerful tool in the environmental field and will lead to the creation of truly portable devices.

  17. Capillaries for use in a multiplexed capillary electrophoresis system

    SciTech Connect

    Yeung, Edward S.; Chang, Huan-Tsang; Fung, Eliza N.

    1997-12-09

    The invention provides a side-entry optical excitation geometry for use in a multiplexed capillary electrophoresis system. A charge-injection device is optically coupled to capillaries in the array such that the interior of a capillary is imaged onto only one pixel. In Sanger-type 4-label DNA sequencing reactions, nucleotide identification ("base calling") is improved by using two long-pass filters to split fluorescence emission into two emission channels. A binary poly(ethyleneoxide) matrix is used in the electrophoretic separations.

  18. Polyacrylamide Gel Electrophoresis for Purification of Large Amounts of RNA.

    PubMed

    Meyer, Mélanie; Masquida, Benoît

    2016-01-01

    Polyacrylamide gel electrophoresis (PAGE) constitutes a powerful technique for the efficient purification of RNA molecules dedicated to applications that require high purity levels. PAGE allows for the fractionation of RNA obtained from cell extracts, chemical or enzymatic synthesis, or modification experiments. Native or denaturing conditions can be chosen for analytical or preparative-scale separations and the nucleotide resolution can be tuned by changing the percentage and reticulation of the gel material. In this protocol, we focus on the preparation of milligram-scale amounts of ~200 nucleotides (nt) RNA molecules that were used in subsequent crystallization experiments. PMID:26227037

  19. Determination of benzylpenicillin in pharmaceuticals by capillary zone electrophoresis

    SciTech Connect

    Hoyt, A.M. Jr. ); Sepaniak, M.J. )

    1989-04-01

    A rapid and direct method is described for the determination of benzylpenicillin (penicillin G) in pharmaceutical preparations. The method involves very little sample preparation and total analysis time for duplicate results is less 30 minutes per sample. The method takes advantage of the speed and separating power of capillary zone electrophoresis (CZE). Detection of penicillin is by absorption at 228 nm. An internal standard is employed to reduce sample injection error. The method was applied successfully to both tablets and injectable preparations. 14 refs., 5 figs., 3 tabs.

  20. The fluid mechanics of continuous flow electrophoresis in perspective

    NASA Technical Reports Server (NTRS)

    Saville, D. A.

    1980-01-01

    Buoyancy alters the flow in continuous flow electrophoresis chambers through the mechanism of hydrodynamic instability and, when the instability is supressed by careful cooling of the chamber boundaries, by restructuring the axial flow. The expanded roles of buoyancy follow upon adapting the size of the chamber and the electric field so as to fractionate certain sorts of cell populations. Scale-up problems, hydrodynamic stability and the altered flow fields are discussed to show how phenomena overlooked in the design and operations of narrow-gap devices take on an overwhelming importance in wide-gap chambers

  1. Human muscle proteins: analysis by two-dimensional electrophoresis

    SciTech Connect

    Giometti, C.S.; Danon, M.J.; Anderson, N.G.

    1983-09-01

    Proteins from single frozen sections of human muscle were separated by two-dimensional gel electrophoresis and detected by fluorography or Coomassie Blue staining. The major proteins were identical in different normal muscles obtained from either sex at different ages, and in Duchenne and myotonic dystrophy samples. Congenital myopathy denervation atrophy, polymyositis, and Becker's muscular dystrophy samples, however, showed abnormal myosin light chain compositions, some with a decrease of fast-fiber myosin light chains and others with a decrease of slow-fiber light chains. These protein alterations did not correlate with any specific disease, and may be cause by generalized muscle-fiber damage.

  2. Analysis of mutations using PCR and denaturing gradient gel electrophoresis

    SciTech Connect

    Cariello, N.F.; Swenberg, J.A. Duke Univ., Durham, NC ); DeBellis, A.; Skopek, T.R. )

    1991-01-01

    Denaturing gradient gel electrophoresis (DGGE) separates DNA molecules based on primary sequence. Under the appropriate conditions, all base pair (bp) substitutions, frameshifts, and deletions less than about 10 bp can be resolved from the wild type sequence using DGGE. Polymerase chain reaction (PCR) permits facile amplification of a given region of the genome. The authors have combined PCR and DGGE to: (1) localize mutations in the X-linked human androgen receptor gene; (2) analyze thousands of thioguanine-resistant mutants simultaneously; (3) examine the fidelity of several DNA polymerases used in PCR.

  3. Two-dimensional gel electrophoresis data in support of leaf comparative proteomics of two citrus species differing in boron-tolerance.

    PubMed

    Sang, Wen; Huang, Zeng-Rong; Qi, Yi-Ping; Yang, Lin-Tong; Guo, Peng; Chen, Li-Song

    2015-09-01

    Here, we provide the data from a comparative proteomics approach used to investigate the response of boron (B)-tolerant 'Xuegan' (Citrus sinensis) and B-intolerant 'Sour pummelo' (Citrus grandis) leaves to B-toxicity. Using two-dimensional gel electrophoresis (2-DE) technique, we identified 50 and 45 protein species with a fold change of more than 1.5 and a P-value of less than 0.05 from B-toxic C. sinensis and C. grandis leaves. These B-toxicity-responsive protein species were mainly involved in carbohydrate and energy metabolism, antioxidation and detoxification, stress responses, coenzyme biosynthesis, protein and amino acid metabolism, signal transduction, cell transport, cytoskeleton, nucleotide metabolism, and cell cycle and DNA processing. A detailed analysis of this data may be obtained from Sang et al. (J. Proteomics 114 (2015))[1]. PMID:26217760

  4. A study of cell electrophoresis as a means of purifying growth hormone secreting cells

    NASA Technical Reports Server (NTRS)

    Plank, Lindsay D.; Hymer, W. C.; Kunze, M. Elaine; Marks, Gary M.; Lanham, J. Wayne

    1983-01-01

    Growth hormone secreting cells of the rat anterior pituitary are heavily laden with granules of growth hormone and can be partialy purified on the basis of their resulting high density. Two methods of preparative cell electrophoresis were investigated as methods of enhancing the purification of growth hormone producing cells: density gradient electrophoresis and continuous flow electrophoresis. Both methods provided a two- to four-fold enrichment in growth hormone production per cell relative to that achieved by previous methods. Measurements of electrophoretic mobilities by two analytical methods, microscopic electrophoresis and laser-tracking electrophoresis, revealed very little distinction between unpurified anterior pituitary cell suspensions and somatotroph-enriched cell suspensions. Predictions calculated on the basis of analytical electrophoretic data are consistent with the hypothesis that sedimentation plays a significant role in both types of preparative electrophoresis and the electrophoretic mobility of the growth hormone secreting subpopulation of cells remains unknown.

  5. Monitoring refolding of tailspike endorhamnosidase using capillary electrophoresis-laser induced tryptophan fluorescence

    SciTech Connect

    Jensen, P.K.; Lee, Cheng S.; King, J.A.

    1997-12-31

    The use of capillary electrophoresis equipped with laser-induced tryptophan fluorescence detection is presented for monitoring the refolding pathway of phage P22 tailspike endorhamnosidase. Upon initiation of refolding, tailspike polypeptides rapidly fold into structured monomeric intermediates with a high content of secondary structure. These monomeric species associate to form the triple-chain defined folding intermediates, the protrimers. Conversion of the protrimer into the native, sodium dodecyl sulfate (SDS) resistant tailspike protein is the rate-limiting step in the refolding pathway. Refolding kinetics and yield measured by capillary electrophoresis are in good agreement with those obtained via native gel electrophoresis, SDS polyacrylamide gel electrophoresis (SDS-PAGE) and fluorescence spectrophotometry. To enhance separation resolution between protrimer and native protein in capillary electrophoresis, the use of poly(ethylene oxide) is investigated for the introduction of a sieving separation mechanism. The increased viscosity of the electrophoresis buffer may also play a role in resolution enhancement.

  6. BioMEMS and Electrophoresis in 2006: Review of the 23rd Annual Meeting of the American Electrophoresis Society

    PubMed Central

    Minerick, Adrienne R.; Ugaz, Victor M.

    2007-01-01

    The 23rd Annual Meeting of the American Electrophoresis Society (AES) was held at the San Francisco Hilton in San Francisco, California on 12–17 November 2006. This year’s meeting featured a look toward the future, with an emphasis on theoretical and experimental advances in miniaturization of BioMEMS, electrokinetics, and proteomics technologies. A total of 13 sessions accommodating 71 presentations and 18 posters were held in conjunction with the Annual Meeting of the American Institute of Chemical Engineers (AIChE). This review and corresponding special issue of Biomicrofluidics provide a sampling of some of the exciting research presented at the conference. PMID:19693377

  7. Startup of electrophoresis in a suspension of colloidal spheres.

    PubMed

    Chiang, Chia C; Keh, Huan J

    2015-12-01

    The transient electrophoretic response of a homogeneous suspension of spherical particles to the step application of an electric field is analyzed. The electric double layer encompassing each particle is assumed to be thin but finite, and the effect of dynamic electroosmosis within it is incorporated. The momentum equation for the fluid outside the double layers is solved through the use of a unit cell model. Closed-form formulas for the time-evolving electrophoretic and settling velocities of the particles in the Laplace transform are obtained in terms of the electrokinetic radius, relative mass density, and volume fraction of the particles. The time scale for the development of electrophoresis and sedimentation is significantly smaller for a suspension with a higher particle volume fraction or a smaller particle-to-fluid density ratio, and the electrophoretic mobility at any instant increases with an increase in the electrokinetic particle radius. The transient electrophoretic mobility is a decreasing function of the particle volume fraction if the particle-to-fluid density ratio is relatively small, but it may increase with an increase in the particle volume fraction if this density ratio is relatively large. The particle interaction effect in a suspension on the transient electrophoresis is much weaker than that on the transient sedimentation of the particles. PMID:26417706

  8. Simple and robust monitoring of ethanol fermentations by capillary electrophoresis.

    PubMed

    Oliver, James D; Sutton, Adam T; Karu, Naama; Phillips, Michael; Markham, Julie; Peiris, Paul; Hilder, Emily F; Castignolles, Patrice

    2015-01-01

    Free-solution capillary electrophoresis (CE), or capillary zone electrophoresis, with direct UV detection was used for the first time for the determination of mono- and disaccharides, sugar alcohols, and ethanol in fermentation broths. Sample preparation proved to be minimal: no derivatization or specific sample purification was needed. The CE conditions can be adapted to the type of fermentation by simply altering the background electrolyte (BGE). KOH (130 mM) or NaOH (130 mM) as the BGE led to the fastest analysis time when monitoring simple fermentations. A mixture of 65 mM NaOH and 65 mM LiOH led to a 19% improvement in resolution for a complex mixture of carbohydrates. Quantification of a simple carbohydrate fermentation by CE showed values in close agreement with that of high-performance anion exchange chromatography and high-performance liquid chromatography (HPLC) on a cation exchange resin. For complex fermentations, quantification of carbohydrates by HPLC and CE led to similar results, whereas CE requires an injection volume of only 10-20 nL. Analysis of an ethanol fermentation of hydrolyzed plant fiber demonstrated the robustness of the separation and detection of carbohydrates, as well as ethanol. Ethanol determination is achieved by coupling the CE method to pressure mobilization, using the same instrument and the same sample. PMID:25040822

  9. Chip-based capillary electrophoresis with an electrodeless nanospray interface.

    PubMed

    Vrouwe, E X; Gysler, J; Tjaden, U R; van der Greef, J

    2000-01-01

    A sheathless and electrodeless nanospray interface has been used to interface a polycarbonate capillary electrophoresis (CE) chip to a mass spectrometer (MS). The chip was made of two flat polycarbonate plates which were bolted together. Channels were imprinted in one of the plates with metal wires, using a hydraulic press. A short tapered capillary connected to the chip was used as the nanospray emitter. The advantage of this electrodeless interface is that it was not necessary to apply a electrospray voltage to the chip or the nanospray emitter. Instead, the CE voltage already applied to the buffer compartment on the chip, to drive the electrophoresis, was used to generate the spray also. A low conductivity buffer of 1.25 mmol/L ammonium acetate in 80% methanol was used to obtain a large electric field across the buffer channel. The performance of the device was evaluated by analyzing a mixture of three beta-agonists Relative standard deviation (RSD) values obtained were between 4.8 and 5.0%. A sample concentration of 40 nmol/L resulted in a signal-to-noise ratio of 2 to 5 for the different components. Compared to a conventional CE analysis in a fused silica capillary with UV detection, only a minor loss of resolution was observed, which can be attributed to the design of the chip. PMID:10962491

  10. Separation of Trivalent Actinides from Lanthanides Using a Capillary Electrophoresis

    SciTech Connect

    Mori, Tomotaka; Ishii, Yasuo; Hayashi, Kazunori; Suganuma, Hideo; Satoh, Isamu

    2007-07-01

    A separation of {sup 241}Am(III) from {sup 152,154}Eu(III) was carried out using a capillary electrophoresis technique in a mixed solvent (CH{sub 3}OH/H{sub 2}O) system containing thiocyanate ion. First, the formation constants ({beta}{sub n}) between thiocyanate ion and Eu(III) or Am(III) were investigated in the mixed solvent solutions by a back-extraction technique using bis (2-ethylhexyl) hydrogen phosphate-toluene. The mean charges calculated on the basis of the data of {beta}{sub n} for Eu(III) were comparatively higher than those for Am(III). Based on the differences between the mean charges of Eu(III) and Am(III), separations for Am(III)/Eu(III) by means of capillary electrophoresis technique were tried in the (H{sup +}, Na{sup +})(SCN{sup -}, ClO{sub 4}{sup -}) mixed solvent solutions. It was proved that Am(III) was completely separated from Eu(III). (authors)

  11. Development of coatings to control electroosmosis in zero gravity electrophoresis

    NASA Technical Reports Server (NTRS)

    Krupnick, A. C.

    1974-01-01

    A major problem confronting the operation of free fluid electrophoresis in zero gravity is the control of electrokinetic phenomena and, in particular, electroosmosis. Due to the severity of counter flow, as a result of electroosmosis, the electrical potential developed at the surface of shear must be maintained at near, or as close to, zero millivolts as possible. Based upon this investigation, it has been found that the amount of bound water or the degree of hydroxylation plays a major role in the control of this phenomena. Of necessity, factors, such as adhesion, biocompatibility, protein adsorption, and insolubility were considered in this investigation because of the long buffer-coating exposure times required by present space operations. Based upon tests employing microcapillary electrophoresis, it has been found that gamma amino propyl trihydroxysilane produced a coating which provides the lowest potential (minus 3.86 mv) at the surface of shear between the stationary and mobile layers. This coating has been soaked in both borate and saline buffers, up to three months, in a pH range of 6.5 to 10 without deleterious effects or a change in its ability to control electrokinetic effects.

  12. The trajectories of spheres during agarose gel electrophoresis.

    PubMed

    Griess, G A; Harris, R A; Serwer, P

    1993-01-01

    To develop a physical description of the gel-induced retardation of spheres during gel electrophoresis, the microscopic motion of single electrically charged latex spheres is statistically quantified here, by digital image analysis. To obtain adequate resolution in space, comparatively large spheres, 240 nm in radius, are used. The following observations are made during electrophoresis in a 0.2% agarose gel at 22 degrees C: (a) When a comparatively high field (3.0 V cm-1) is used, inelastic collisions result in field-induced trapping of spheres; no elastic collisions are observed. (b) Reduction of the field from 3.0 to 0.0 V cm-1 results in reverse migration of previously trapped spheres. (c) In the absence of trapping, the electrical field does not cause an alteration in the tortuosity of motion (i.e. motion in a field-perpendicular direction). (d) When results are obtained for a constant time between images (0.2 s), gel-dependent deviations from a true random walk are not observed in the absence of trapping. (e) When results are obtained as a function of time between images, significant gel-dependent deviation from a random walk is observed. In the absence of trapping, the data presented here indicate that retardation is derived primarily from dissipative processes that are concentrated near gel fibers. However, steric effects have not yet been distinguished from hydrodynamic effects. PMID:8199223

  13. Separation of radiolabelled glycosaminoglycan oligosaccharides by polyacrylamide-gel electrophoresis.

    PubMed Central

    Hampson, I N; Gallagher, J T

    1984-01-01

    Glycosaminoglycan oligosaccharides generated by treatment of biosynthetically radiolabelled dermatan sulphate and hyaluronic acid with chondroitin AC lyase or testicular hyaluronidase may be resolved into a series of discrete bands by polyacrylamide-gel electrophoresis. Bands were identified by fixation in glacial acetic acid containing 20% (w/v) 2,5-diphenyloxazole followed by fluorography. The bands represented glycans which differed in size by one disaccharide unit. For the larger oligosaccharides (decasaccharides and above) of similar charge: mass ratio, there was a linear relationship between electrophoretic mobility and log Mr. However, the smaller species showed anomalous migration patterns. Consideration of the structures of the fragments produced by the different enzyme treatments suggests that copolymeric and homopolymeric oligosaccharides may be separated by polyacrylamide-gel electrophoresis. There are many potential applications of this technique, foremost amongst them being studies on the molecular size heterogeneity and patterns of enzyme-mediated depolymerization of native glycosaminoglycan chains and investigations into rates of polymer chain elongation and post-polymerization modification reactions so essential to glycosaminoglycan function. Images Fig. 1. Fig. 2. Fig. 5. Fig. 6. PMID:6477495

  14. An enhanced capillary electrophoresis method for characterizing natural organic matter.

    PubMed

    Cottrell, Barbara A; Cheng, Wei Ran; Lam, Buuan; Cooper, William J; Simpson, Andre J

    2013-02-21

    Natural organic matter (NOM) is ubiquitous and is one of the most complex naturally occurring mixtures. NOM plays an essential role in the global carbon cycle; atmospheric and natural water photochemistry; and the long-range transport of trace compounds and contaminants. There is a dearth of separation techniques capable of resolving this highly complex mixture. To our knowledge, this is the first reported use of ultrahigh resolution counterbalance capillary electrophoresis to resolve natural organic matter. The new separation strategy uses a low pH, high concentration phosphate buffer to reduce the capillary electroosmotic flow (EOF). Changing the polarity of the electrodes reverses the EOF to counterbalance the electrophoretic mobility. Sample stacking further improves the counterbalance separation. The combination of these conditions results in an electropherogram comprised up to three hundred peaks superimposed on the characteristic "humic hump" of NOM. Fraction collection, followed by three-dimensional emission excitation spectroscopy (EEMs) and UV spectroscopy generated a distinct profile of fluorescent and UV absorbing components. This enhanced counterbalance capillary electrophoresis method is a potentially powerful technique for the characterization and separation of NOM and complex environmental mixtures in general. PMID:23289095

  15. Capillary electrophoresis of small ssDNA molecules

    NASA Astrophysics Data System (ADS)

    Kopecka, Katerina; Slater, Gary W.; Drouin, Guy

    2004-03-01

    Recently, the electrophoretic separation of small ssDNA fragments (bellow 250 bases) has attracted a lot of attention because of applications related to Single Nucleotide Polymorphisms. In order to optimize these systems, we require a better understanding of DNA migration behavior in this size range. While the reptation model provides an excellent understanding of the dynamics of long DNA fragments in gel electrophoresis, the properties of small DNA fragments has not been studied extensively yet. At least three theoretical formulas have been proposed to explain the mobility of short ssDNA molecules in this regime. Specifically, the Ogston regime was introduced for small molecules having radii-of-gyration comparable to or smaller than the pore size of the sieving matrix. We introduce these three different formulas and discuss how their free parameters are related to actual physical parameters. We then test these formulas with new data obtained by capillary electrophoresis in our laboratory using poly(dimethylacrylamide) sieving matrices. Our results show that all three formulas provide decent fits, and that their fitting parameters are consistent with one another. This is the first step towards the development of a systematic approach to optimizing sequencing systems for this size range.

  16. Increasing Transport Efficiencies of Polymer Based Solar Cells by Electrophoresis

    NASA Astrophysics Data System (ADS)

    Wong, Terrence

    2011-03-01

    Organic polymer photovoltaic (PV) cells are an active area of Applied Physics research because of four unique characteristics: (1) relatively inexpensive costs, (2) transparent properties, (3) flexibility, and (4) ease of mass production. We are studying the effects of incorporating single-walled carbon nanotubes (SWCNs) into a mixture of poly-(3-hexylthiophene) (P3HT), to test the affects on transport characteristics. The experiment will be segregated into parallel trials, with fixed volume ratios of P3HT:SWCNs to test the effects of (1) random orientation of SWCNs or the control, and (2) an aligned orientation of SWCNs. An electrophoresis-based technique, similar to gel electrophoresis, used to separate DNA fragments of variable masses, is used for partial alignment of the SWCN. Fixed geometry metalized substrates in a four striped copper patternare used for the transport studies and the P3HT:SWCN film's resistivity is monitored in-situ. The oriented films show enhanced conductivity, indicating this plays a major role in the increased efficiencies found in P3HT:SWCN based polymer solar cells.

  17. Increasing Transport Efficiencies of Polymer Based Solar Cells by Electrophoresis

    NASA Astrophysics Data System (ADS)

    Wong, Terrence

    2010-10-01

    Organic polymer photovoltaic (PV) cells are an active area of Applied Physics research because of four unique characteristics: (1) relatively inexpensive costs, (2) transparent properties, (3) flexibility, and (4) ease of mass production. We are studying the effects of incorporating single-walled carbon nanotubes (SWCNs) into a mixture of poly-(3-hexylthiophene) (P3HT), to test the affects on transport characteristics. The experiment will be segregated into parallel trials, with fixed volume ratios of P3HT:SWCNs to test the effects of (1) random orientation of SWCNs or the control, and (2) an aligned orientation of SWCNs. An electrophoresis-based technique, similar to gel electrophoresis, used to separate DNA fragments of variable masses, is used for partial alignment of the SWCN. Fixed geometry metalized substrates in a four striped copper patternare used for the transport studies and the P3HT:SWCN film's resistivity is monitored in-situ. The oriented films show enhanced conductivity, indicating this plays a major role in the increased efficiencies found in P3HT:SWCN based polymer solar cells.

  18. Studies on proteinograms in dermatorphytes by disc electrophoresis. Part 2: Protein bands of keratinophilic fungi

    NASA Technical Reports Server (NTRS)

    Danev, P.; Balabanov, V.; Friedrich, E.

    1983-01-01

    Disc electrophoresis studies on keratinophili fungi demonstrated corresponding proteinograms in morphologically homogeneous strains of the same species, but different in different species of one and the same genus.

  19. Proteomic analysis of surface proteins of Trichinella spiralis muscle larvae by two-dimensional gel electrophoresis and mass spectrometry

    PubMed Central

    2013-01-01

    Background Trichinella spiralis is a zoonotic tissue-dwelling parasitic nematode that infects humans and other mammals. Its surface proteins are recognized as antigenic in many infected hosts, being directly exposed to the host’s immune system and are the main target antigens that induce the immune responses. The larval surface proteins may also interact with intestinal epithelial cells and may play an important role in the invasion and development process of T. spiralis. The purpose of this study was to analyze and characterize the surface proteins of T. spiralis muscle larvae by two-dimensional gel electrophoresis (2-DE) and mass spectrometry. Methods The surface proteins of T. spiralis muscle larvae were stripped from the cuticle of live larvae by the cetyltrimethylammonium bromide (CTAB) and sodium deoxycholate. The surface protein stripping was examined by an immunofluorescent test (IFT). The surface proteins were analyzed by SDS-PAGE and Western blotting, and then identified by 2-DE and MALDI-TOF/TOF mass spectrometry analysis. Results The IFT results showed that the surface proteins-stripped larvae were not recognized by sera of mice immunized with surface antigens. Western blotting showed 7 of 12 protein bands of the surface proteins were recognized by mouse infection sera at 18 dpi and at 42 dpi. The 2-DE results showed that a total of approximately 33 proteins spots were detected with molecular weights varying from 10 to 66 kDa and isoelectric point (pI) from 4 to 7. Twenty-seven of 33 protein spots were identified and characterized to correlate with 15 different proteins. Out of the 14 proteins identified as T. spiralis proteins, 5 proteins (partial P49 antigen, deoxyribonuclease II family protein, two serine proteases, and serine proteinase) had catalytic and hydrolase activity. All of these 5 proteins were also associated with metabolic processes and 2 of the five proteins were associated with cellular processes. Conclusions In this study, T

  20. A new 2-dimensional method for constructing visualized treatment objectives for distraction osteogenesis of the short mandible.

    PubMed

    van Beek, H

    2010-01-01

    Open bite development during distraction of the mandible is common and partly due to inaccurate planning of the treatment. Conflicting guidelines exist in the literature. A method for Visualized Treatment Objective (VTO) construction is presented as an aid for determining the correct orientation of monodirectional and multidirectional distractors. Distraction on the left and on the right side of the mandible takes place in a parallel manner in order to maintain intercondylar width. It follows that in the absence of marked asymmetry, the amount of mandibular body distraction, the amount of ramus distraction and (should it apply), the amount of closure of the gonial angle, can be derived from a simple 2-dimensional plan. After presurgical orthodontic treatment, a cephalogram is taken and a VTO is constructed, that aims at a good occlusion with the enhanced mandible in centric relation, with little or no change of the original position of the rami. PMID:19837600

  1. Impact of mode partition noise in free-running gain-switched Fabry-Perot laser for 2-dimensional OCDMA.

    PubMed

    Wang, Xu; Chan, Kam

    2004-07-26

    Free-running gain-switched Fabry-Perot laser diode is an appropriate incoherent broadband optical source for incoherent 2-dimensional optical code division multiple access. However, the mode partition noise (MPN) in the laser seriously degrades performance. We derived a bit error rate (BER) expression in the presence of MPN using the power spectra of the laser. The theory agreed with the experimental results. There was a power penalty and BER floor due to the MPN in the laser. Therefore, this scheme should be operated with a sufficiently large number of modes. At least 9 modes should be used for error-free transmission at 1 Gbit/s for the laser we investigated in this work. PMID:19483858

  2. Effect of Cardiac Resynchronization Therapy on Left Atrial Size and Function as Expressed by Speckle Tracking 2-Dimensional Strain.

    PubMed

    Valzania, Cinzia; Gadler, Fredrik; Boriani, Giuseppe; Rapezzi, Claudio; Eriksson, Maria J

    2016-07-15

    Changes in left atrial (LA) strain in patients treated with cardiac resynchronization therapy (CRT) remain not entirely explored. We prospectively evaluated long-term changes in LA size and function and their relation with left ventricular (LV) reverse remodeling and noninvasive hemodynamic variables in patients treated with CRT by 2-dimensional speckle tracking echocardiography. Thirty patients (62 ± 11 years, 63% men) underwent 2-dimensional speckle tracking echocardiography before implant and after 12 months. LA area, global and regional LA strains, LV ejection fraction (LVEF) and longitudinal strain, mitral regurgitation (MR), and diastolic variables were evaluated. At 12 months, CRT responders (60%) exhibited an increase in LA strain (11.4 ± 6.5% vs 16.5 ± 7.9%, p <0.001) and a reduction in LA area (p = 0.002), which were associated with an improvement in MR, E/E' ratio, LVEF, and LV longitudinal strain. In nonresponders, a worsening in LA strain (11.4 ± 6.8% vs 8.7 ± 4.6%, p = 0.017) and LA area (p = 0.002) occurred in parallel with an increase in E/E', whereas LVEF and LV longitudinal strain were unchanged. In conclusion, over long-term follow-up, LA size and strain improved in CRT responders, while worsening in nonresponders. Changes in LV function, filling pressures, and MR seem to be related to LA reverse remodeling, giving a feedback loop. PMID:27241837

  3. Buffer optimization for high resolution of human lung cancer tissue proteins by two-dimensional gel electrophoresis.

    PubMed

    Lee, Kibeom; Pi, Kyungbae; Lee, Keeman

    2009-01-01

    A problem in proteomic analysis of lung cancer tissue is the presence of complex components of different histological backgrounds (squamous cell carcinoma, small cell lung carcinoma, and adenocarcinoma). The efficient solubilization of protein components before two-dimensional electrophoresis (2-DE) is a very critical. Poor solubilization has been associated with a failure to detect proteins and diffuse, streaked and/or trailing protein spots. Here, we have optimized the solubilization of human lung cancer tissue to increase protein resolution. Isoelectric focusing (IEF) rehydration buffer containing a thiourea-urea mixture provided superior resolution, whereas a buffer without thiourea yielded consistently poor results. In addition, IEF rehydration buffers containing CHAPS and DTT gave superior resolution, whereas buffers containing Nonidet P-40 (NP-40) and/or Triton X-100 did not. A tributylphosphine-containing buffer gave consistently poor results. Using optimized conditions, we used 2-D gel analysis of human lung cancer tissue to identify 11 differentially-expressed protein spots by MALDI-mass spectrometry. This study provides a methodological tool to study the complex mammalian proteomes. PMID:18800191

  4. Phytotoxin coronatine enhances heat tolerance via maintaining photosynthetic performance in wheat based on Electrophoresis and TOF-MS analysis

    PubMed Central

    Zhou, Yuyi; Zhang, Mingcai; Li, Jianmin; Li, Zhaohu; Tian, Xiaoli; Duan, Liusheng

    2015-01-01

    Coronatine (COR) is a phytotoxin produced by Pseudomonas syringae. Its structure is similar to Jasmonates, which play a number of diverse roles in plant defense. Both have the COI1 plant receptor, so coronatine can manipulate plant hormone signaling to access nutrients and counteract defense responses. In addition to the hormone system, coronatine affects plant nitrogenous metabolism and chloroplast ultrastructure. In this study, we first examined a typical nitrogen-losing phenotype, and used the polyacrylamide gel approach to demonstrate soluble total protein patterns in a time-course experiment under different temperature conditions. We then employed dimensional gel electrophoresis technology (2-DE) and MALDI-TOF-MS to sequester and identify the sensitive proteins. We found a total of 27 coronatine sensitive proteins, 22 of which were located in the chloroplast and 6 of which were directly involved in photosynthesis. Finally, we measured levels of chlorophyll and photosynthetic performance to reveal the phenotypic effect of these proteins. Taken together, these results demonstrated that coronatine enhanced heat tolerance by regulating nitrogenous metabolism and chloroplast ultrastructure to maintain photosynthetic performance and reduce yield loss under heat stress. PMID:26347991

  5. Proteomic Analysis of Rice Nonhost Resistance to Puccinia striiformis f. sp. tritici Using Two-Dimensional Electrophoresis

    PubMed Central

    Zhao, Jing; Yang, Yuheng; Kang, Zhensheng

    2014-01-01

    Rice (Oryza sativa L.) is the only widely cultivated gramineous crops that cannot be infected by rust fungi. To decipher the molecular basis of rice nonhost resistance (NHR) to Puccinia striiformis f. sp. tritici (Pst), the causal agent of wheat stripe rust, proteomic analysis was performed using the two-dimensional electrophoresis (2-DE) technique. The expressed proteins from rice leaves 24 and 48 h post inoculation with Pst and from mock-inoculated leaves were identified. Quantitative analysis revealed a total of 27 differentially expressed proteins in response to Pst inoculation. Most of these proteins fall into the category “response to stimulus” and are involved in basic resistance processes, such as glycerol-3-phosphate and hydrogen peroxide signaling. A homologue of wheat leaf rust resistance protein Lr10 was also identified, implicating multiple layers of plant defense are implicated in rice NHR to Pst. These results demonstrate an intrinsic relationship between host and nonhost resistance. Changes in abundance of these proteins, together with their putative functions reveal a comprehensive profile of rice NHR to Pst and provide new insights into plant immunity. PMID:25429427

  6. Capillary zone electrophoresis and capillary electrophoresis-mass spectrometry for analyzing qualitative and quantitative variations in therapeutic albumin.

    PubMed

    Marie, Anne-Lise; Przybylski, Cédric; Gonnet, Florence; Daniel, Régis; Urbain, Rémi; Chevreux, Guillaume; Jorieux, Sylvie; Taverna, Myriam

    2013-10-24

    The present study describes a reproducible and quantitative capillary zone electrophoresis (CZE) method, which leads to the separation of nine forms (native, oxidized and glycated) of human serum albumin (HSA). In an attempt to identify the different species separated by this CZE method, the capillary electrophoresis was coupled to mass spectrometry using a sheath liquid interface, an optimized capillary coating and a suitable CE running buffer. CE-MS analyses confirmed the heterogeneity of albumin preparation and revealed new truncated and modified forms such as Advanced Glycation End products (AGEs). Assignment of the CZE peaks was carried out using specific antibodies, carboxypeptidase A or sample reduction before or during the CE separation. Thus, five HSA forms were unambiguously identified. Using this CZE method several albumin batches produced by slightly different fractionation ways could be discriminated. Furthermore, analyses of HSA preparations marketed by five pharmaceutical industries revealed that two therapeutic albumins, including that marketed by LFB, contained the highest proportion of native form and lower levels of oxidized forms. PMID:24120174

  7. Effect of separation dimensions on resolution and throughput using very narrow-range IEF for 2-DE after solution phase isoelectric fractionation of a complex proteome.

    PubMed

    Lee, KiBeom; Pi, KyungBae

    2009-04-01

    This paper describes how the lengths of the first and second dimensions in narrow pH-range 2-DE affect the number of detected protein spots, by analysis of human breast carcinoma cell line lysates prefractionated by solution phase IEF. The aim is to maximize throughput while minimizing experimental costs. In this study, systematic evaluation of narrow-range IPG strip lengths showed that separation distances were very important, with dramatic increases in resolution when longer gels were used. Compared with 7 cm minigels, maximal resolution was obtained using 18 and 24 cm IPG strips. Systematic evaluation of SDS-PAGE gel length showed a far weaker influence of separation length on resolution in the second dimension compared with that observed for the IEF dimension. There was little benefit in using separation distances greater than 12-15 cm, at least with currently available electrophoresis units. The work shows that regions of the IPG strip not containing any proteins can be excised to fit a smaller gel if prefractionation using IEF in solution has been performed. As expected, larger 2-DE gel volumes resulting from the use of longer IPG strips and second dimension gels decreased detection sensitivity when equal protein loads were used. However, this effect could be readily eliminated by increasing the loads applied to IPG strips. PMID:19301322

  8. Evaluation of two-dimensional electrophoresis and liquid chromatography – tandem mass spectrometry for tissue-specific protein profiling of laser-microdissected plant samples

    SciTech Connect

    Schad, Martina; Lipton, Mary S.; Giavalisco, Patrick; Smith, Richard D.; Kehr, Julia

    2005-07-14

    Laser microdissection (LM) allows the collection of homogeneous tissue- and cell specific plant samples. The employment of this technique with subsequent protein analysis has thus far not been reported for plant tissues, probably due to the difficulties associated with defining a reasonable cellular morphology and, in parallel, allowing efficient protein extraction from tissue samples. The relatively large sample amount needed for successful proteome analysis is an additional issue that complicates protein profiling on a tissue- or even cell-specific level. In contrast to transcript profiling that can be performed from very small sample amounts due to efficient amplification strategies, there is as yet no amplification procedure for proteins available. In the current study, we compared different tissue preparation techniques prior to LM/laser pressure catapulting (LMPC) with respect to their suitability for protein retrieval. Cryosectioning was identified as the best compromise between tissue morphology and effective protein extraction. After collection of vascular bundles from Arabidopsis thaliana stem tissue by LMPC, proteins were extracted and subjected to protein analysis, either by classical two-dimensional gel electrophoresis (2-DE), or by high-efficiency liquid chromatography (LC) in conjunction with tandem mass spectrometry (MS/MS). Our results demonstrate that both methods can be used with LMPC collected plant material. But because of the significantly lower sample amount required for LC-MS/MS than for 2-DE, the combination of LMPC and LC-MS/MS has a higher potential to promote comprehensive proteome analysis of specific plant tissues.

  9. [Detection of picobirnaviruses by electrophoresis of RNA in polyacrylamide gel].

    PubMed

    Novikova, N A; Epifanova, N V; Fedorova, O F; Golitsyna, L N; Kupriianova, N V

    2003-01-01

    Double-segment profiles typical of picobirnavirus (PBV) were detected (during 1994-2001) in nucleic acids extracted from feces of children (3 cases) and calf (1 case) with diarrhea by using the method of electrophoresis. The human genomic PBV segments migrated in the polyacrylamide gel (PAAG) as dsRNA segments sized 1.7 and 2.4 kbp for small and large segments, respectively; the similar calf sizes were 1.5 and 2.6 kbp. PBVs were detected in various places of the Nizhny Novgorod Region and at different time periods. It was for the first time that the PBV circulation was proven to be present in Russia's territory. However, their association with diarrhea was not reliably established, and the pathogenic PBV potential needs further investigations. PMID:14708231

  10. Protein separation by continuous-flow electrophoresis in microgravity.

    PubMed

    Clifton, M J; Roux-de Balmann, H; Sanchez, V

    1996-07-01

    During the IML-2 space shuttle mission, the RAMSES instrument was operated in the Spacelab module. This continuous-flow electrophoresis device performs separation and purification of protein solutions on a preparative scale. Samples containing artificial mixtures of pure proteins were used to test the capabilities of the device, and useful separations were obtained for proteins having a mobility difference of only 3 x 10(-9) m2 V-1 s-1. Operating conditions that cannot be applied on earth were explored for two different sample concentrations, one of which is too high to allow treatment on earth. It agrees well with a previously published numerical model in that the main cause of loss in resolution in this process is the electrohydrodynamic spreading of the protein filaments. PMID:11539848

  11. Separability of electrostatic and hydrodynamic forces in particle electrophoresis

    NASA Astrophysics Data System (ADS)

    Todd, Brian A.; Cohen, Joel A.

    2011-09-01

    By use of optical tweezers we explicitly measure the electrostatic and hydrodynamic forces that determine the electrophoretic mobility of a charged colloidal particle. We test the ansatz of O'Brien and White [J. Chem. Soc. Faraday IIJCFTBS0300-923810.1039/f29787401607 74, 1607 (1978)] that the electrostatically and hydrodynamically coupled electrophoresis problem is separable into two simpler problems: (1) a particle held fixed in an applied electric field with no flow field and (2) a particle held fixed in a flow field with no applied electric field. For a system in the Helmholtz-Smoluchowski and Debye-Hückel regimes, we find that the electrostatic and hydrodynamic forces measured independently accurately predict the electrophoretic mobility within our measurement precision of 7%; the O'Brien and White ansatz holds under the conditions of our experiment.

  12. Active colloids propelled by induced-charge electrophoresis

    NASA Astrophysics Data System (ADS)

    Han, Ming; Luijten, Erik

    Populations of motile organisms exhibit a variety of collective behaviors, ranging from bacterial colony formation to the flocking of birds. Current understanding of these active motions, which are typically far from equilibrium and based on the collective behavior of self-propelled entities, is far from complete. One approach is to reproduce these observations in systems of synthetic active colloids. However, one of the standard self-propulsion mechanisms, induced-charge electrophoresis (ICEP) of a dielectric Janus colloid remains not fully understood by itself, especially the strong dependence of the resultant particle motion on the frequency of the external field. Resolution of this outstanding problem requires detailed study of the time-resolved dielectric response of the colloid and the dynamics of the electric double layer. Through molecular dynamics simulations coupled with an efficient dielectric solver, we elucidate the underlying mechanism of the frequency dependence of ICEP and the polarization of a metallodielectric Janus colloid.

  13. Thermally reversible gels in electrophoresis. I - Matrix characterization

    NASA Technical Reports Server (NTRS)

    Righetti, Pier Giorgio; Snyder, Robert S.

    1988-01-01

    Two series of thermally reversible hydrogen-bonded gels have been characterized: (5 pct) PVA-(4 pct) PEG and (5 pct) PVA-(0.04 pct) borate gels. They both have extremely low melting points (16-17 C) and could be of potential interest for recovery of proteins after preparative electrophoresis. The PVA-borate gels can be exploited in the pH range 7-11 by progressively increasing the borate content in the pH interval 8 to 7 and concomitantly decreasing the borate levels in the pH zone 8 to 11. It is hypothesized that the low melting point of these gels is due to the fact that they are sparingly and sparsely hydrogen bonded along the PVA chain: on the average, 1 OH group out of 3 or 4 OH groups in the PVA polymer should be engaged in H-bond formation.

  14. Separation and detection of DNA by capillary electrophoresis.

    PubMed

    Kasper, T J; Melera, M; Gozel, P; Brownlee, R G

    1988-12-23

    The use of capillary electrophoresis for the separation and detection of nucleic acids has been investigated. Lab-model instruments have been built, using commercially available UV absorbance and fluorescence detectors which were modified for use with 50-100 microns I.D. fused-silica capillary tubing. The sensitivity of these instruments (signal-to-noise ratio = 3) was measured as 15 micrograms/ml for fluorescence detection of ethidium bromide-stained herring-sperm DNA and 3 micrograms/ml for UV absorbance detection. With the former instruments a variety of strategies has been used to attain rapid separations of bases, oligonucleotides, restriction fragments and whole phage, viral and plasmid DNAs. PMID:3266219

  15. Isolating aptamers using capillary electrophoresis-SELEX (CE-SELEX).

    PubMed

    Mosing, Renee K; Bowser, Michael T

    2009-01-01

    SELEX (systematic evolution of ligands by exponential enrichment) is a process for isolating DNA or RNA sequences with high affinity and selectivity for molecular targets from random sequence libraries. These sequences are commonly referred to as aptamers. The process typically requires 10-15 cycles of enrichment, PCR amplification and nucleic acid purification to obtain high-affinity aptamers. We have demonstrated that using capillary electrophoresis (CE) as an enrichment step greatly improves the efficiency of the process. CE-SELEX is capable of isolating high-affinity aptamers in as little as 2-4 rounds of selection, shortening the process time from several weeks to as little as a few days. PMID:19377982

  16. Moving towards harmonized reporting of serum and urine protein electrophoresis.

    PubMed

    Moss, Michael A

    2016-06-01

    During the last decade, surveys by questionnaire in Canada, Australia and New Zealand revealed wide variation in reporting practices by laboratories and individual practitioners in the interpretation of serum and urine protein electrophoresis (PE). Such variation has potential to adversely impact patient outcomes if report structure is inconsistent or if the messaging is incorrectly perceived by the receiving physician. Concerted efforts have been initiated to promote harmonization in the use of interpretative comments. The primary goal is to add value through clear communication with requesting physicians in the interest of quality patient care. Resistance to a harmonized approach largely reflects longstanding personal reporting habits and preferences but change can be more readily embraced if the new system is intuitive, easy to use and saves time in reporting. PMID:26824981

  17. Separation of ions in acidic solution by capillary electrophoresis

    SciTech Connect

    Thornton, M.

    1997-10-08

    Capillary electrophoresis (CE) is an effective method for separating ionic species according to differences in their electrophoretic mobilities. CE separations of amino acids by direct detection are difficult due to their similar electrophoretic mobilities and low absorbances. However, native amino acids can be separated by CE as cations at a low pH by adding an alkanesulfonic acid to the electrolyte carrier which imparts selectivity to the system. Derivatization is unnecessary when direct UV detection is used at 185 nm. Simultaneous speciation of metal cations such as vanadium (IV) and vanadium (V) can easily be performed without complexation prior to analysis. An indirect UV detection scheme for acidic conditions was also developed using guanidine as the background carrier electrolyte (BCE) for the indirect detection of metal cations. Three chapters have been removed for separate processing. This report contains introductory material, references, and general conclusions. 80 refs.

  18. Protein Separation by Capillary Gel Electrophoresis: A Review

    PubMed Central

    Zhu, Zaifang; Lu, Joann J.; Liu, Shaorong

    2011-01-01

    Capillary gel electrophoresis (CGE) has been used for protein separation for more than two decades. Due to the technology advancement, current CGE methods are becoming more and more robust and reliable for protein analysis, and some of the methods have been routinely used for the analysis of protein-based pharmaceuticals and quality controls. In light of this progress, we survey 147 papers related to CGE separations of proteins and present an overview of this technology. We first introduce briefly the early development of CGE. We then review the methodology, in which we specifically describe the matrices, coatings, and detection strategies used in CGE. CGE using microfabricated channels and incorporation of CGE with two-dimensional protein separations are also discussed in this section. We finally present a few representative applications of CGE for separating proteins in real-world samples. PMID:22122927

  19. Cyclodextrins in capillary electrophoresis: recent developments and new trends.

    PubMed

    Escuder-Gilabert, L; Martín-Biosca, Y; Medina-Hernández, M J; Sagrado, S

    2014-08-29

    Despite the fact that extensive research in the field of separations by capillary electrophoresis (CE) has been carried out and many reviews have been published in the last years, a specific review on the use and future potential of cyclodextrins (CDs) in CE is not available. This review focuses the attention in the CD-CE topic over the January 2013-February 2014 period (not covered by previous more general CE-reviews). Recent contributions (reviews and research articles) including practical uses (e.g. solute-CD binding constant estimation and further potentials; 19% of publications), developments and applications (mainly chiral and achiral analysis; 38 and 24% of publications, respectively) are summarized in nine comprehensive tables and are commented. Statistics and predictions related to the CD-CE publications are highlighted in order to infer the current and expected research interests. Finally, trends and initiatives on CD-CE attending to real needs or practical criteria are outlined. PMID:24947884

  20. Accessing Protein Methyltransferase and Demethylase Enzymology Using Microfluidic Capillary Electrophoresis

    PubMed Central

    Wigle, Tim J.; Provencher, Laurel M.; Norris, Jacqueline L.; Jin, Jian; Brown, Peter J.; Frye, Stephen V.; Janzen, William P.

    2010-01-01

    Summary The discovery of small molecules targeting the > 80 enzymes that add (methyltransferases) or remove (demethylases) methyl marks from lysine and arginine residues, most notably present in histone tails, may yield unprecedented chemotherapeutic agents and facilitate regenerative medicine. To better enable chemical exploration of these proteins, we have developed a novel and highly quantitative microfluidic capillary electrophoresis assay to enable full mechanistic studies of these enzymes and the kinetics of their inhibition. This technology separates small biomolecules, i.e., peptides, based on their charge-to-mass ratio. Methylation, however, does not alter the charge of peptide substrates. To overcome this limitation, we have employed a methylation-sensitive endoproteinase strategy to separate methylated from unmethylated peptides. The assay was validated on a lysine methyltransferase (G9a) and a lysine demethylase (LSD1) and was employed to investigate the inhibition of G9a by small molecules. PMID:20659682

  1. Theoretical and experimental separation dynamics in capillary zone electrophoresis

    NASA Technical Reports Server (NTRS)

    Thormann, Wolfgang; Michaud, Jon-Pierre; Mosher, Richard A.

    1986-01-01

    The mathematical model of Bier et al. (1983) is used in a computer aided analysis of the conditions in capillary zone electrophoresis (ZE) under which sample zones migrate noninteractively with the carrier electrolyte. The monitoring of sample zones with a capillary analyzer that features both on-line conductivity and UV detection at the end of the separation trough is discussed. Data from a ZE analysis of a 5-component mixture are presented, and it is noted that all five components can be monitored via their conductivity change if enough sample is present. It is suggested from the results that the concentration ratio of background buffer to sample should be a minimum of 100:1 to effectively apply the plate concept to ZE.

  2. Ultrasensitive nucleic acid sequence detection by single-molecule electrophoresis

    SciTech Connect

    Castro, A; Shera, E.B.

    1996-09-01

    This is the final report of a one-year laboratory-directed research and development project at Los Alamos National Laboratory. There has been considerable interest in the development of very sensitive clinical diagnostic techniques over the last few years. Many pathogenic agents are often present in extremely small concentrations in clinical samples, especially at the initial stages of infection, making their detection very difficult. This project sought to develop a new technique for the detection and accurate quantification of specific bacterial and viral nucleic acid sequences in clinical samples. The scheme involved the use of novel hybridization probes for the detection of nucleic acids combined with our recently developed technique of single-molecule electrophoresis. This project is directly relevant to the DOE`s Defense Programs strategic directions in the area of biological warfare counter-proliferation.

  3. Modified electrokinetic sample injection method in chromatography and electrophoresis analysis

    DOEpatents

    Davidson, J. Courtney; Balch, Joseph W.

    2001-01-01

    A sample injection method for horizontal configured multiple chromatography or electrophoresis units, each containing a number of separation/analysis channels, that enables efficient introduction of analyte samples. This method for loading when taken in conjunction with horizontal microchannels allows much reduced sample volumes and a means of sample stacking to greatly reduce the concentration of the sample. This reduction in the amount of sample can lead to great cost savings in sample preparation, particularly in massively parallel applications such as DNA sequencing. The essence of this method is in preparation of the input of the separation channel, the physical sample introduction, and subsequent removal of excess material. By this method, sample volumes of 100 nanoliter to 2 microliters have been used successfully, compared to the typical 5 microliters of sample required by the prior separation/analysis method.

  4. Recent advances in affinity capillary electrophoresis for binding studies.

    PubMed

    Albishri, Hassan M; El Deeb, Sami; AlGarabli, Noura; AlAstal, Raghda; Alhazmi, Hassan A; Nachbar, Markus; El-Hady, Deia Abd; Wätzig, Hermann

    2014-01-01

    The present review covers recent advances and important applications of affinity capillary electrophoresis (ACE). It provides an overview about various ACE types, including ACE-MS, the multiple injection mode, the use of microchips and field-amplified sample injection-ACE. The most common scenarios of the studied affinity interactions are protein-drug, protein-metal ion, protein-protein, protein-DNA, protein-carbohydrate, carbohydrate-drug, peptide-peptide, DNA-drug and antigen-antibody. Approaches for the improvements of ACE in term of precision, rinsing protocols and sensitivity are discussed. The combined use of computer simulation programs to support data evaluation is presented. In conclusion, the performance of ACE is compared with other techniques such as equilibrium dialysis, parallel artificial membrane permeability assay, high-performance affinity chromatography as well as surface plasmon resonance, ultraviolet, circular dichroism, nuclear magnetic resonance, Fourier transform infrared, fluorescence, MS and isothermal titration calorimetry. PMID:25534793

  5. Bio-nano interactions detected by nanochannel electrophoresis.

    PubMed

    Luan, Binquan

    2016-08-01

    Engineered nanoparticles have been widely used in industry and are present in many consumer products. However, their bio-safeties especially in a long term are largely unknown. Here, a nanochannel-electrophoresis-based method is proposed for detecting the potential bio-nano interactions that may further lead to damages to human health and/or biological environment. Through proof-of-concept molecular dynamics simulations, it was demonstrated that the transport of a protein-nanoparticle complex is very different from that of a protein along. By monitoring the change of ionic currents induced by a transported analyte as well as the transport velocities of the analyte, the complex (with bio-nano interaction) can be clearly distinguished from the protein alone (with no interaction with tested nanoparticles). PMID:27334561

  6. Microchip capillary electrophoresis based electroanalysis of triazine herbicides.

    PubMed

    Islam, Kamrul; Chand, Rohit; Han, Dawoon; Kim, Yong-Sang

    2015-01-01

    The number of pesticides used in agriculture is increasing steadily, leading to contamination of soil and drinking water. Herein, we present a microfluidic platform to detect the extent of contamination in soil samples. A microchip capillary electrophoresis system with in-channel electrodes was fabricated for label-free electroanalytical detection of triazine herbicides. The sample mixture contained three representative triazines: simazine, atrazine and ametryn. The electropherogram for each individual injection of simazine, atrazine and ametryn showed peaks at 58, 66 and 72 s whereas a mixture of them showed distinct peaks at 59, 67 and 71 s respectively. The technique as such may prove to be a useful qualitative and quantitative tool for the similar environmental pollutants. PMID:25231112

  7. Electrophoresis of semiflexible heteropolymers and the ``hydrodynamic Kuhn length''

    NASA Astrophysics Data System (ADS)

    Chubynsky, Mykyta V.; Slater, Gary W.

    Semiflexible polymers, such as DNA, are rodlike for short lengths and coil-like for long lengths. For purely geometric properties, such as the end-to-end distance, the crossover between these two behaviors occurs when the polymer length is on the order of the Kuhn length. On the other hand, for the hydrodynamic friction coefficient it is easy to see by comparing the expressions for a rod and a coil that the crossover should occur at the polymer length, termed by us the hydrodynamic Kuhn length, which is larger than the ordinary Kuhn length by a logarithmic factor that can be quite significant. We show that for the problem of electrophoresis of a heteropolymer consisting of several blocks of (in general) different stiffnesses, both of these length scales can be important depending on the details of the problem.

  8. Simian Virus 40 Deoxyribonucleic Acid Synthesis: Analysis by Gel Electrophoresis

    PubMed Central

    Tegtmeyer, Peter; Macasaet, Francisco

    1972-01-01

    An agarose-gel electrophoresis technique has been developed to study simian virus 40 deoxyribonucleic acid (DNA) synthesis. Superhelical DNA I, relaxed DNA II, and replicative intermediate (RI) molecules were clearly resolved from one another for analytical purposes. Moreover, the RI molecules could be identified as early or late forms on the basis of their electrophoretic migration in relation to that of DNA II. The technique has been utilized to study the kinetics of simian virus 40 DNA synthesis in pulse and in pulse-chase experiments. The average time required to complete the replication of prelabeled RI molecules and to convert them into DNA I was approximately 10 min under the experimental conditions employed. PMID:4343542

  9. Electrophoresis of DNA on a disordered two-dimensional substrate

    NASA Astrophysics Data System (ADS)

    Reichhardt, C. J. Olson; Reichhardt, C.

    2006-11-01

    We propose a method for electrophoretic separation of DNA in which adsorbed polymers are driven over a disordered two-dimensional substrate which contains attractive sites for the polymers. Using simulations of a model for long polymer chains, we show that the mobility increases with polymer length, in contrast to gel electrophoresis techniques, and that separation can be achieved for a range of length scales. We demonstrate that the separation mechanism relies on steric interactions between polymer segments, which prevent substrate disorder sites from trapping more than one DNA segment each. Since thermal activation does not play a significant role in determining the polymer mobility, band broadening due to diffusion can be avoided in our separation method.

  10. Electrophoresis of DNA on a disordered two-dimensional substrate

    NASA Astrophysics Data System (ADS)

    Olson Reichhardt, Cynthia J.; Reichhardt, Charles

    2007-03-01

    We propose a method for electrophoretic separation of DNA in which adsorbed polymers are driven over a disordered two-dimensional substrate which contains attractive sites for the polymers. Using simulations of a model for long polymer chains, we show that the mobility increases with polymer length, in contrast to gel electrophoresis techniques, and that separation can be achieved for a range of length scales. We demonstrate that the separation relies on steric interactions between polymer segments, which prevent substrate disorder sites from trapping more than one DNA segment each. Since thermal activation does not play a significant role in determining the polymer mobility, band broadening due to diffusion can be avoided in our separation method. [1] Phys. Rev. E 74, 051908 (2006).

  11. Molecular Dynamics Simulation of Electrophoresis of a Telehelic Polymer Chain

    NASA Astrophysics Data System (ADS)

    Bhattacharya, Aniket

    2002-08-01

    We report the conformational and the dynamical properties of an end-labeled polymer chain embedded in a porous medium made of randomly distributed immobile spherical obstacles using a stochastic Molecular Dynamic(MD) simulation method for several obstacle densities and for various bias strenghts applied only to one end of the chain. First, various properties of the chain are studied when the external bias is set to zero. We then extend the stochastic MD simulation to study the electrophresis of a polymer chain driven by (i) a steady and (ii) a time dependent electric field. These studies are relevant for various time dependent gel electrophoresis methods widely used to separate DNA molecules. The qualitative features are compared with experiments, analytic theories, and recent Monte Carlo Simulation results.

  12. Nanomaterial surface chemistry design for advancements in capillary electrophoresis modes.

    PubMed

    Ivanov, Michael R; Haes, Amanda J

    2011-01-01

    Tailored surface chemistry impacts nanomaterial function and stability in applications including in various capillary electrophoresis (CE) modes. Although colloidal nanoparticles were first integrated as colouring agents in artwork and pottery over 2000 years ago, recent developments in nanoparticle synthesis and surface modification increased their usefulness and incorporation in separation science. For instance, precise control of surface chemistry is critically important in modulating nanoparticle functionality and stability in dynamic environments. Herein, recent developments in nanomaterial pseudostationary and stationary phases will be summarized. First, nanomaterial core and surface chemistry compositions will be classified. Next, characterization methods will be described and related to nanomaterial function in various CE modes. Third, methods and implications of nanomaterial incorporation into CE will be discussed. Finally, nanoparticle-specific mechanisms likely involved in CE will be related to nanomaterial surface chemistry. Better understanding of surface chemistry will improve nanoparticle design for the integration into separation techniques. PMID:20967383

  13. Preparation of MgB2 superconducting tapes using electrophoresis

    NASA Astrophysics Data System (ADS)

    Xu, J. D.; Wang, S. F.; Zhou, Y. B.; Zhou, Y. L.; Chen, Z. H.; Cui, D. F.; Lu, H. B.; He, M.; Dai, S. Y.; Yang, G. Z.

    2002-08-01

    Superconducting MgB2/Ta tapes with a critical temperature of 34 K have been prepared successfully by ex situ annealing of electrophoresis-grown boron in the presence of Mg vapour at 920 °C. Scanning electron microscopy was used to examine the surface morphology of the MgB2/Ta tapes, and well-formed MgB2 crystals with sizes up to 2 μm were observed. The x-ray diffraction patterns showed randomly orientated growth of MgB2 phase in the tapes. Estimates using hysteresis loops and the Bean model give a value of 6.8 × 105 A cm-2 for the critical current density.

  14. Potentials and Method Improvements of Capillary Zone Electrophoresis for Use in Spelt Breeding Programs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Capillary zone electrophoresis (CZE) in acidic buffer systems is capable of separating cereal storage proteins based on similar separation principles as classical acidic polyacrylamide gel electrophoresis. However, it is faster, its resolution is distinctly higher and data evaluation is much simpler...

  15. Design, development, test, and evaluation of an automated analytical electrophoresis apparatus

    NASA Technical Reports Server (NTRS)

    Bartels, P. A.; Bier, M.

    1977-01-01

    An Automated Analytical Electrophoresis Apparatus (AAEA) was designed, developed, assembled, and preliminarily tested. The AAEA was demonstrated to be a feasible apparatus for automatically acquiring, displaying, and storing (and eventually analyzing) electrophoresis mobility data from living blood cells. The apparatus and the operation of its major assemblies are described in detail.

  16. Instrumental development of novel detection and separation methods for capillary electrophoresis

    SciTech Connect

    Garner, T.

    1993-07-01

    After a general introduction, this thesis is divided into 3 parts: indirect fluorescence detection of sugars separated by capillary zone electrophoresis with visible laser excitation, absorption detection in capillary electrophoresis by fluorescence energy transfer, and increased selectivity for electrochromatography by dynamic ion exchange.

  17. Analysis of Anions in Ambient Aerosols by Microchip Capillary Electrophoresis

    SciTech Connect

    Liu, Yan; MacDonald, David A.; Yu, Xiao-Ying; Hering, Susanne V.; Collett, Jeffrey L.; Henry, Charles S.

    2006-10-01

    We describe a microchip capillary electrophoresis method for the analysis of nitrate and sulfate in ambient aerosols. Investigating the chemical composition of ambient aerosol particles is essential for understanding their sources and effects. Significant progress has been made towards developing mass spectrometry-based instrumentation for rapid qualitative analysis of aerosols. Alternative methods for rapid quantification of selected high abundance compounds are needed to augment the capacity for widespread routine analysis. Such methods could provide much higher temporal and spatial resolution than can be achieved currently. Inorganic anions comprise a large percentage of particulate mass with nitrate and sulfate among the most abundant species. While ion chromatography has proven very useful for analyzing extracts of time-integrated ambient aerosol samples collected on filters and for semi-continuous, on-line particle composition measurements, there is a growing need for development of new compact, inexpensive approaches to routine on-line aerosol ion analysis for deployment in spatially dense, atmospheric measurement networks. Microchip capillary electrophoresis provides the necessary speed and portability to address this need. In this report, on-column contact conductivity detection is used with hydrodynamic injection to create a simple microchip instrument for analysis of nitrate and sulfate. On-column contact conductivity detection was achieved using a Pd decoupler placed upstream from the working electrodes. Microchips containing two Au or Pd working electrodes showed a good linear range (5-500 µM) and low limits-of-detection for sulfate and nitrate with Au providing the lowest detection limits (1 µM) for both ions. The completed microchip system was used to analyze ambient aerosol filter samples. Nitrate and sulfate concentrations measured by the microchip matched the concentrations measured by ion chromatography.

  18. Gene analysis of multiple oral bacteria by the polymerase chain reaction coupled with capillary polymer electrophoresis.

    PubMed

    Liu, Chenchen; Yamaguchi, Yoshinori; Sekine, Shinichi; Ni, Yi; Li, Zhenqing; Zhu, Xifang; Dou, Xiaoming

    2016-03-01

    Capillary polymer electrophoresis is identified as a promising technology for the analysis of DNA from bacteria, virus and cell samples. In this paper, we propose an innovative capillary polymer electrophoresis protocol for the quantification of polymerase chain reaction products. The internal standard method was modified and applied to capillary polymer electrophoresis. The precision of our modified internal standard protocol was evaluated by measuring the relative standard deviation of intermediate capillary polymer electrophoresis experiments. Results showed that the relative standard deviation was reduced from 12.4-15.1 to 0.6-2.3%. Linear regression tests were also implemented to validate our protocol. The modified internal standard method showed good linearity and robust properties. Finally, the ease of our method was illustrated by analyzing a real clinical oral sample using a one-run capillary polymer electrophoresis experiment. PMID:26648455

  19. Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat

    PubMed Central

    2010-01-01

    Background Low-molecular-weight glutenin subunits (LMW-GS) play a crucial role in determining end-use quality of common wheat by influencing the viscoelastic properties of dough. Four different methods - sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE, IEF × SDS-PAGE), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and polymerase chain reaction (PCR), were used to characterize the LMW-GS composition in 103 cultivars from 12 countries. Results At the Glu-A3 locus, all seven alleles could be reliably identified by 2-DE and PCR. However, the alleles Glu-A3e and Glu-A3d could not be routinely distinguished from Glu-A3f and Glu-A3g, respectively, based on SDS-PAGE, and the allele Glu-A3a could not be differentiated from Glu-A3c by MALDI-TOF-MS. At the Glu-B3 locus, alleles Glu-B3a, Glu-B3b, Glu-B3c, Glu-B3g, Glu-B3h and Glu-B3j could be clearly identified by all four methods, whereas Glu-B3ab, Glu-B3ac, Glu-B3ad could only be identified by the 2-DE method. At the Glu-D3 locus, allelic identification was problematic for the electrophoresis based methods and PCR. MALDI-TOF-MS has the potential to reliably identify the Glu-D3 alleles. Conclusions PCR is the simplest, most accurate, lowest cost, and therefore recommended method for identification of Glu-A3 and Glu-B3 alleles in breeding programs. A combination of methods was required to identify certain alleles, and would be especially useful when characterizing new alleles. A standard set of 30 cultivars for use in future studies was chosen to represent all LMW-GS allelic variants in the collection. Among them, Chinese Spring, Opata 85, Seri 82 and Pavon 76 were recommended as a core set for use in SDS-PAGE gels. Glu-D3c and Glu-D3e are the same allele. Two new alleles, namely, Glu-D3m in cultivar Darius, and Glu-D3n in Fengmai 27, were identified by 2-DE. Utilization of the suggested standard cultivar set, seed of

  20. Carrier-number fluctuations in the 2-dimensional electron gas at the LaAlO{sub 3}/SrTiO{sub 3} interface

    SciTech Connect

    Barone, C. Romeo, F.; Pagano, S.; Di Gennaro, E.; Miletto Granozio, F.; Scotti di Uccio, U.; Pallecchi, I.; Marrè, D.

    2013-12-02

    The voltage-spectral density S{sub V} (f) of the 2-dimensional electron gas formed at the interface of LaAlO{sub 3}/SrTiO{sub 3} has been thoroughly investigated. The low-frequency component has a clear 1/f behavior with a quadratic bias current dependence, attributed to resistance fluctuations. However, its temperature dependence is inconsistent with the classical Hooge model, based on carrier-mobility fluctuations. The experimental results are, instead, explained in terms of carrier-number fluctuations, due to an excitation-trapping mechanism of the 2-dimensional electron gas.

  1. Carrier-number fluctuations in the 2-dimensional electron gas at the LaAlO3/SrTiO3 interface

    NASA Astrophysics Data System (ADS)

    Barone, C.; Romeo, F.; Pagano, S.; Di Gennaro, E.; Miletto Granozio, F.; Pallecchi, I.; Marrè, D.; Scotti di Uccio, U.

    2013-12-01

    The voltage-spectral density SV (f) of the 2-dimensional electron gas formed at the interface of LaAlO3/SrTiO3 has been thoroughly investigated. The low-frequency component has a clear 1/f behavior with a quadratic bias current dependence, attributed to resistance fluctuations. However, its temperature dependence is inconsistent with the classical Hooge model, based on carrier-mobility fluctuations. The experimental results are, instead, explained in terms of carrier-number fluctuations, due to an excitation-trapping mechanism of the 2-dimensional electron gas.

  2. Optimal Charging Profiles with Minimal Intercalation-Induced Stresses for Lithium-Ion Batteries Using Reformulated Pseudo 2-Dimensional Models

    SciTech Connect

    Suthar, B; Northrop, PWC; Braatz, RD; Subramanian, VR

    2014-07-30

    This paper illustrates the application of dynamic optimization in obtaining the optimal current profile for charging a lithium-ion battery by restricting the intercalation-induced stresses to a pre-determined limit estimated using a pseudo 2-dimensional (P2D). model. This paper focuses on the problem of maximizing the charge stored in a given time while restricting capacity fade due to intercalation-induced stresses. Conventional charging profiles for lithium-ion batteries (e.g., constant current followed by constant voltage or CC-CV) are not derived by considering capacity fade mechanisms, which are not only inefficient in terms of life-time usage of the batteries but are also slower by not taking into account the changing dynamics of the system. (C) The Author(s) 2014. Published by ECS. This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives 4.0 License (CC BY-NC-ND, http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is not changed in any way and is properly cited. For permission for commercial reuse, please email: oa@electrochem.org. All rights reserved.

  3. Phase change memory devices formed by using 2 dimensional layered Graphene-In2 Se3 van der Waals heterostructure

    NASA Astrophysics Data System (ADS)

    Choi, Min Sup; Yang, Chenxi; Ra, Chang Ho; Yoo, Won Jong

    Indium selenide (In2Se3) is one of the unique materials which have both a layered structure and phase change property. One of the advantages of using 2 dimensional (2D) materials is their potential to form van der Waals heterostructures which enable unique physical properties and novel quantum device functions, which cannot be achieved in 2D material alone. In this study, we fabricated vertically stacked graphene-In2Se3 heterostructured memory devices. The fabricated devices showed a rapid increase of current conduction, which is attributed to the phase transition of In2Se3. The TEM images demonstrated that In2Se3 transformed from polycrystalline to layered structure thanks to the effective thermal confinement effect between graphene and In2Se3, attributed to the low thermal conductivity of layered materials in vertical direction. In addition, the current conduction could be controlled effectively by applying different pulse voltages, showing stable retention and endurance characteristics. It is thought that the differently bonded states contribute to this control process. This study demonstrates the possibility of Graphene-In2Se3 van der Waals heterostructure as 2D based future memory electronics. This work was supported by the National Research Foundation of Korea(NRF) Grant funded by the Korea government(MEST) (No. 2013R1A2A2A01015516).

  4. Separation of biogenic materials by electrophoresis under zero gravity (L-3)

    NASA Technical Reports Server (NTRS)

    Kuroda, Masao

    1993-01-01

    Electrophoresis separates electrically charged materials by imposing a voltage between electrodes. Though free-flow electrophoresis is used without carriers such as colloids to separate and purify biogenic materials including biogenic cells and proteins in blood, its resolving power and separation efficiency is very low on Earth due to sedimentation, flotation, and thermal convection caused by the specific gravity differences between separated materials and buffer solutions. The objective of this experiment is to make a comparative study of various electrophoresis conditions on the ground and in zero-gravity in order to ultimately develop a method for separating various important 'vial' components which are difficult to separate on the ground.

  5. [Determination of glyoxalate and oxalate by capillary zone electrophoresis].

    PubMed

    Guan, Jin; Wang, Huize; Ren, Liyan; Niu, Qiuling

    2012-01-01

    A method for the simultaneous determination of glyoxalate and oxalate by capillary zone electrophoresis (CZE) was developed. The influences of type, concentration and pH of the running buffer, and the applied voltage on separation were investigated. Glyoxalate and oxalate were separated within 11 min under the conditions of 20 mmol/L borax-5.5 mmol/L potassium hydrogen phthalate (pH 9.0), applied voltage of 20 kV, and detected wavelength of 212 nm. The calibration curves of glyoxalate and oxalate showed good linearity in the ranges of 0.8 -20 g/L and 1.2-20 g/L, respectively. The correlation coefficients were 0.999 3 and 0.997 5, respectively. The limits of detection for glyoxalate and oxalate were 0.2 and 0.4 g/L (S/N = 3), respectively. The average recoveries at three spiked levels were 98.3%-102.5% with acceptable relative standard deviations of 0.35%-0.61%. This method is simple, low cost and high performance. The method was successfully used for the determination of glyoxalate and oxalate in real samples, and the assay results were satisfactory. PMID:22667103

  6. Miniaturized movable contactless conductivity detection cell for capillary electrophoresis.

    PubMed

    Macka, Miroslav; Hutchinson, Joseph; Zemann, Andreas; Shusheng, Zhang; Haddad, Paul R

    2003-06-01

    A miniaturized capacitively coupled contactless conductivity detector (mini-C(4)D) cell has been designed which is small enough to allow it to slide along the effective capillary length inside the capillary cassette of an Agilent capiillary electrophoresis system (CE) (or other CE brand of similar construction), including the possibility of positioning it close to the point of optical detection (4 cm), or even putting two such detector cells in one cassette. The cell was tested and the performance characteristics (noise, sensitivity, and peak width) were compared with those obtained with the previously used large C(4)D cell. No significant differences were observed. The mini-C(4)D was used in simultaneous separations of common cations and anions where its advantage over a larger C(4)D cell is the ability to vary the point of detection with the mini-C(4)D cell continuously at any point along the capillary length, so that the optimum apparent selectivity can be chosen. Other applications include providing a convenient second point of detection in addition to photometric detection, such as to measure accurately the linear velocity of a zone, or to allow placement of two mini-C(4)D cells in one capillary cassette simultaneously. PMID:12858387

  7. Travelling Wave Electrophoresis Due To Selective Currents At Electrodes

    NASA Astrophysics Data System (ADS)

    Booth, William; Edwards, Boyd

    2015-03-01

    Abstract Using COMSOL finite element modeling software we simulate a 2D traveling-wave electrophoresis (TWE) device for microfluidic chromatography and electrolyte concentration. A periodic array of four electrodes each produce AC potentials shifted by a quarter-period from one another. This yields an electric wave which travels down the channel. Ions of varying mobilities in solution are carried along with the electric wave or left behind at different rates. We employ a simplified model for asymmetric reactions at the electrodes in order to solve the issue of electric double layer shielding at the electrodes. The selective reactions allow for the formation of diffusion layers of ions which attempt to follow the traveling electric wave. We examine the formation of these diffusion layers and how various system parameters affect motion of various ions through the system. With easy control over the traveling electric wave's frequency and direction one may employ this method for concentrating or separating bands of electrolytes. NSF Grant 1066730.

  8. Quantification of sugars in breakfast cereals using capillary electrophoresis.

    PubMed

    Toutounji, Michelle R; Van Leeuwen, Matthew P; Oliver, James D; Shrestha, Ashok K; Castignolles, Patrice; Gaborieau, Marianne

    2015-05-18

    About 80% of the Australian population consumes breakfast cereal (BC) at least five days a week. With high prevalence rates of obesity and other diet-related diseases, improved methods for monitoring sugar levels in breakfast cereals would be useful in nutrition research. The heterogeneity of the complex matrix of BCs can make carbohydrate analysis challenging or necessitate tedious sample preparation leading to potential sugar loss or starch degradation into sugars. A recently established, simple and robust free solution capillary electrophoresis (CE) method was used in a new application to 13 BCs (in Australia) and compared with several established methods for quantification of carbohydrates. Carbohydrates identified in BCs by CE included sucrose, maltose, glucose and fructose. The CE method is simple requiring no sample preparation or derivatization and carbohydrates are detected by direct UV detection. CE was shown to be a more robust and accurate method for measuring carbohydrates than Fehling method, DNS (3,5-dinitrosalicylic acid) assay and HPLC (high performance liquid chromatography). PMID:25841355

  9. Xanthine derivatives quantification in serum by capillary zone electrophoresis.

    PubMed

    Peris-Vicente, Juan; Rambla-Alegre, Maria; Durgavanshi, Abhilasha; Bose, Devasish; Esteve-Romero, Josep; Marco-Peiró, Sergio

    2014-10-01

    A capillary electrophoresis method was developed to quantify caffeine and theophylline, xanthine derivatives with bronchodilator activity. Buffer concentration, pH and applied voltage were optimized using a central composite design-face centred. Separation conditions were: silica capillary tube, 75 μm (i.d.) and 61 cm (total length); absorbance detection, 280 nm; borate buffer, 20 mM, pH 9.0; applied voltage, 25 kV and 1 psi injection/8 s. Validation was performed in blank serum following the International Conference Harmonization guidelines: resolution (peaks without overlapping), linear range (0.125-50 µg/mL; r(2) > 0.9999), limits of detection and quantification (10; 20 and 33; 66 ppb for caffeine and theophylline, respectively), intra- and inter-day precision (Relative standard deviation lower than 1.9%) and accuracy (98-101%). Migration times were <8 min. This method is simple, specific and suitable and reaches high label claims (98.7-100.4%) in pharmaceutical formulations analysis. Moreover, the method was applied to the monitoring of the analytes in serum of patients. PMID:24220991

  10. Study on Dicarboxylic Acids in Aerosol Samples with Capillary Electrophoresis

    PubMed Central

    Adler, Heidi; Sirén, Heli

    2014-01-01

    The research was performed to study the simultaneous detection of a homologous series of α, ω-dicarboxylic acids (C2–C10), oxalic, malonic, succinic, glutaric, adipic, pimelic, suberic, azelaic, and sebacic acids, with capillary electrophoresis using indirect UV detection. Good separation efficiency in 2,6-pyridinedicarboxylic acid as background electrolyte modified with myristyl trimethyl ammonium bromide was obtained. The dicarboxylic acids were ionised and separated within five minutes. For the study, authentic samples were collected onto dry cellulose membrane filters of a cascade impactor (12 stages) from outdoor spring aerosols in an urban area. Hot water and ultrasonication extraction methods were used to isolate the acids from membrane filters. Due to the low concentrations of acids in the aerosols, the extracts were concentrated with solid-phase extraction (SPE) before determination. The enrichment of the carboxylic acids was between 86 and 134% with sample pretreatment followed by 100-time increase by preparation of the sample to 50 μL. Inaccuracy was optimised for all the sample processing steps. The aerosols contained dicarboxylic acids C2–C10. Then, mostly they contained C2, C5, and C10. Only one sample contained succinic acid. In the study, the concentrations of the acids in aerosols were lower than 10 ng/m3. PMID:24729915

  11. Combining Capillary Electrophoresis with Mass Spectrometry for Applications in Proteomics

    SciTech Connect

    Simpson, David C.; Smith, Richard D.

    2005-04-01

    Throughout the field of global proteomics, ranging from simple organism studies to human medical applications, the high sample complexity creates demands for improved separations and analysis techniques. Furthermore, with increased organism complexity, the correlation between proteome and genome becomes less certain due to extensive mRNA processing prior to translation. In this way, the same DNA sequence can potentially code for regions in a number of distinct proteins; quantitative differences in expression (or abundance) between these often-related species are of significant interest. Well-established proteomics techniques, which use genomic information to identify peptides that originate from protease digestion, often cannot easily distinguish between such gene products; intact protein-level analyses are required to complete the picture, particularly for identifying post-translational modifications. While chromatographic techniques are currently better suited to peptide analysis, capillary electrophoresis (CE) in combination with mass spectrometry (MS) may become important for intact protein analysis. This review focuses on CE/MS instrumentation and techniques showing promise for such applications, highlighting those with greatest potential. Reference will also be made to developments relevant to peptide-level analyses for use in time- or sample-limited situations.

  12. Capillary electrophoresis separation of the desamino degradation products of oxytocin

    PubMed Central

    Creamer, Jessica S.; Krauss, Shannon T.; Lunte, Susan M.

    2014-01-01

    Oxytocin is an endogenous and therapeutic hormone necessary for maternal health. It is also the subject of fast growing research in the field of behavioral science. This article describes a rapid capillary electrophoresis method using UV detection at 214 nm for the determination of the deamidation products of oxytocin. Deamidation is the most common degradation pathway of peptides and proteins and can lead to reduced therapeutic efficiency of biopharmaceuticals. To achieve a separation of the seven structurally similar desamino peptides from oxytocin, 11 mM sulfobutyl ether β-cyclodextrin and 10% v/v MeOH were added to a background electrolyte of 50 mM phosphate buffer at pH 6.0. The assay is linear within ≤5-100 μM for all species with a total analysis time of 12 min. The method was then applied to monitor the heat-stress degradation of oxytocin at 70°C, where all seven desamino species were observed over a 96 h period. PMID:24166826

  13. Stacking in a continuous sample flow interface in capillary electrophoresis.

    PubMed

    Gstoettenmayr, Daniel; Quirino, Joselito; Ivory, Cornelius F; Breadmore, Michael

    2015-08-21

    Using a tee connector in a commercial capillary electrophoresis instrument, the effect of field amplified sample injection from both flowing and static sample volumes was investigated. It is shown that under identical conditions (40min electrokinetic injection at 5kV from a sample volume of 295μL) the limit of detection using the continuous sample flow interface is 4 times lower than from a static vial. The relationship between different flow rates and injection voltages on the injected sample amount was also investigated using a 2D axisymmetric simulation (COMSOL 4.3b) and verified experimentally, confirming conditions under which there is near-quantitative injection of the sample target ions. Using electrokinetic injection at 30kV and a flow rate of 558nL/s the same enhancement from an even smaller volume of 184μL could be achieved in 5.5min than could be achieved from 295μL and a 40min injection. This sensitivity enhancement factor corresponded to four orders of magnitude improvement compared to a hydrodynamic injection. This is the first report showing that a continuous sample flow interface combined with stacking methods under conditions approaching quantitative injection from the entire sample volume has the potential to be more sensitive than a static system. PMID:26189205

  14. Determination of dissociation constants of cytokinins by capillary zone electrophoresis.

    PubMed

    Barták, P; Bednár, P; Stránský, Z; Bocek, P; Vespalec, R

    2000-05-12

    A method for the pKa determination, based on mobility data measured by capillary zone electrophoresis, was applied to cytokinins and their analogs. The combination of charged mobility standards with an uncharged electroosmosis marker, injected in the uncoated capillary simultaneously with the measured substances, allows one to minimize the number of runs, reduce their duration and, in addition, to inform on the run-to-run stability of electroosmosis and on contingent side-effects. pKa values of investigated cytokinins and their analogs ranged from 2.8 to 4.0 at 25 degrees C in the phosphate and acetate buffers of ionic strength 0.015 M. Standard deviations of the constants, obtained by the non-linear fitting of equations for the pKa calculation, were 3-5-times lower than standard deviations from the linear fitting or from the point-to-point calculation utilizing the Hendersson-Haselbalch equation. The equation of Boltzman sigmoid offers two checks on reliability of effective mobilities that serve as the raw data in the pKa calculation. PMID:10866070

  15. Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

    PubMed

    Bruin, G J; Börnsen, K O; Hüsken, D; Gassmann, E; Widmer, H M; Paulus, A

    1995-08-11

    The approach of using antisense oligonucleotides as potential drugs is based on hybridization of a short chemically-modified oligonucleotide with complementary cellular DNA or RNA sequences. A critical question is the stability of chemically modified antisense oligonucleotides in cellular environments. In a model system, resistance against various nucleases was evaluated by capillary gel electrophoresis (CGE). For some of the samples, matrix assisted laser desorption and ionization mass spectrometry (MALDI-MS) was used as an additional analytical tool to perform stability measurements. Using CGE, the enzymatic degradation of single nucleotides from the oligomer can be followed after different incubation times. 10% T polyacrylamide gels give baseline resolution for oligonucleotides ranging between 5 and 30 bases in length. The kinetic influence of a specific nuclease concentration and the antisense oligonucleotide structure on the cleavage reaction are discussed. Also, a simple desalting method to improve the injection efficiency and sensitivity of the method are described. Examples of measurements of chemically modified antisense 19-mers are presented. PMID:7581844

  16. Electrophoresis-chemiluminescence detection of phenols catalyzed by hemin.

    PubMed

    Shu, Lu; Zhu, Jinkun; Wang, Qingjiang; He, Pingang; Fang, Yuzhi

    2014-09-01

    Based on the catalytic activity of hemin, an efficient biocatalyst, an indirect capillary electrophoresis-chemiluminescence (CE-CL) detection method for phenols using a hemin-luminol-hydrogen peroxide system was developed. Through a series of static injection experiments, hemin was found to perform best in a neutral solution rather than an acidic or alkaline medium. Although halide ions such as Br(-) and F(-) could further enhance the CL signal catalyzed by hemin, it is difficult to apply these conditions to this CE-CL detection system because of the self-polymerization of hemin, as it hinders the CE process. The addition of concentrated ammonium hydroxide to an aqueous/dimethyl sulfoxide solution of hemin-luminol afforded a stable CE-CL baseline. The indirect CE-CL detection of five phenols using this method gave the following limits of detections: 4.8 × 10(-8) mol/L (o-sec-butylphenol), 4.9 × 10(-8) mol/L (o-cresol), 5.4 × 10(-8) mol/L (m-cresol), 5.3 × 10(-8) mol/L (2,4-dichlorophenol) and 7.1 × 10(-8) mol/L (phenol). PMID:24115262

  17. The Application of Pulsed Field Gel Electrophoresis in Clinical Studies

    PubMed Central

    Parizad, Eskandar Gholami; Valizadeh, Azar

    2016-01-01

    Pulsed-field gel electrophoresis is a method applied in separating large segments of deoxyribonucleotide using an alternating and cross field. In a uniform magnetic field, components larger than 50kb pass a route through the gel and since the movement of DNA (Deoxyribonucleic acid) molecules are in a Zigzag form, separation of DNAs as bands carried out better via gel. PFGE in microbiology is a standard method which is used for typing of bacteria. It is also a very useful tool in epidemiological studies and gene mapping in microbes and mammalian cell, also motivated development of large-insert cloning system such as bacterial and yeast artifical chromosomes. In this method, close and similar species in terms of genetic patterns show alike profiles regarding DNA separation, and those ones which don’t have similarity or are less similar, reveal different separation profiles. So this feature can be used to determine the common species as the prevalence agent of a disease. PFGE can be utilized for monitoring and evaluating different micro-organisms in clinical samples and existing ones in soil and water. This method can also be a reliable and standard method in vaccine preparation. In recent decades, PFGE is highly regarded as a powerful tool in control, prevention and monitoring diseases in different populations. PMID:26894068

  18. Electrophoresis system for high temperature mobility measurements of nanosize particles

    NASA Astrophysics Data System (ADS)

    Rodriguez-Santiago, Victor; Fedkin, Mark V.; Lvov, Serguei N.

    2008-09-01

    The electrophoretic mobility, which reflects the zeta potential of a solid material, is an important experimental quantity providing information about the electrical double layer at the solid/liquid interface. A new high temperature electrophoresis cell was developed suitable for electrophoretic mobility measurements of dispersed nanosize particles up to 150 °C and 40 bars. Amorphous silica (SiO2) particle size standards were used to test the particle size detection limit of the new instrument at 25, 100, and 150 °C and several pH values. The microscopic detection of the particles was enabled by dark-field illumination, which allowed extending the previously available capabilities and provided higher accuracy of the electrophoretic mobility data. The electrophoretic mobility measurements for SiO2 at temperatures above 100 °C were reported for the first time and indicated a gradual increase in particle electrophoretic response with increasing temperature. The obtained data indicated negatively charged SiO2 surface throughout the pH and temperature ranges studied.

  19. Theoretical model of electroosmotic flow for capillary zone electrophoresis

    SciTech Connect

    Tavares, M.F.M.; McGuffin, V.L.

    1995-10-15

    A mathematical model of electroosmotic flow in capillary zone electrophoresis has been developed by taking into consideration of the ion-selective properties of silica surfaces. The electroosmotic velocity was experimentally determined, underboth constant voltage and constant current conditions, by using the resistance-monitoring method. A detailed study of electroosmotic flow characteristics in solutions of singly charged, strong electrolytes (NaCl, LiCl, KCl, NaBr, NaI, NaNO{sub 3}, and NaClO{sub 4}), as well as the phosphate buffer system, revealed a linear correlation between the {Zeta} potential and the logarithm of the cation activity. These results suggest that the capillary surface behaves as an ion-selective electrode. Consequently, the {Zeta} potential can be calculated as a function of the composition and pH of the solution with the corresponding modified Nernst equation for ion-selective electrodes. If the viscosity and dielectric constant of the solution are known, the electroosmotic velocity can then be accurately predicted by means of the Helmholtz-Smoluchowski equation. The proposed model has been successfully applied to phosphate buffer solutions in the range of pH from 4 to 10, containing sodium chloride from 5 to 15 mM, resulting in nearly 3% error in the estimation of the electroosmotic velocity. 53 refs., 8 figs., 2 tabs.

  20. Photochemical Microscale Electrophoresis Allows Fast Quantification of Biomolecule Binding.

    PubMed

    Möller, Friederike M; Kieß, Michael; Braun, Dieter

    2016-04-27

    Intricate spatiotemporal patterns emerge when chemical reactions couple to physical transport. We induce electrophoretic transport by a confined photochemical reaction and use it to infer the binding strength of a second, biomolecular binding reaction under physiological conditions. To this end, we use the photoactive compound 2-nitrobenzaldehyde, which releases a proton upon 375 nm irradiation. The charged photoproducts locally perturb electroneutrality due to differential diffusion, giving rise to an electric potential Φ in the 100 μV range on the micrometer scale. Electrophoresis of biomolecules in this field is counterbalanced by back-diffusion within seconds. The biomolecule concentration is measured by fluorescence and settles proportionally to exp(-μ/D Φ). Typically, binding alters either the diffusion coefficient D or the electrophoretic mobility μ. Hence, the local biomolecule fluorescence directly reflects the binding state. A fit to the law of mass action reveals the dissociation constant of the binding reaction. We apply this approach to quantify the binding of the aptamer TBA15 to its protein target human-α-thrombin and to probe the hybridization of DNA. Dissociation constants in the nanomolar regime were determined and match both results in literature and in control experiments using microscale thermophoresis. As our approach is all-optical, isothermal and requires only nanoliter volumes at nanomolar concentrations, it will allow for the fast screening of biomolecule binding in low volume multiwell formats. PMID:27042755

  1. Dielectric Decrement Effects on Nonlinear Electrophoresis of Ideally Polarizable Particles

    NASA Astrophysics Data System (ADS)

    Moran, Jeffrey L.; Chan, Wai Hong Ronald; Buie, Cullen R.; Figliuzzi, Bruno

    2014-11-01

    We present numerical simulations of nonlinear electrophoresis of ideally polarizable particles that specifically include the effects of a spatially non-uniform dielectric permittivity near the particle surface. Models for this dielectric decrement phenomenon have been developed by several authors, including Ben-Yaakov et al. [J. Phys.: Condens. Matter 2009] Hatlo et al. [EPL 2012], and Zhao & Zhai [JFM 2013]. We extend this work to ideally polarizable particles and include the effects of surface conduction and advective transport in the electric double layer. By numerically solving for the coupled velocity field, electric potential, and ionic concentration distributions in the bulk solution surrounding the particle, we demonstrate that the dielectric decrement model predicts ionic saturation around the particle and thus physical implications that resemble those resulting from the steric model developed by Kilic et al. [PRE 2007], albeit with differences that reflect the nonlinearity of the modified Poisson-Boltzmann equation. In addition, we develop a generalized condensed layer model that approximates both the steric and dielectric decrement models in the limits of strong electric fields and negligible surface conduction to obtain more physical insights into these models. We demonstrate that the mobility in both models asymptotically scales as the square root of the electric field at high fields, recovering the result of Bazant et al. [Adv. Colloid Interface Sci. 2009].

  2. Protonation enthalpies of metal oxides from high temperature electrophoresis

    SciTech Connect

    Rodriguez-Santiago, V; Fedkin, Mark V.; Lvov, Serguei N.

    2012-01-01

    Surface protonation reactions play an important role in the behavior of mineral and colloidal systems, specifically in hydrothermal aqueous environments. However, studies addressing the reactions at the solid/liquid interface at temperatures above 100 C are scarce. In this study, newly and previously obtained high temperature electrophoresis data (up to 260 C) zeta potentials and isoelectric points for metal oxides, including SiO2, SnO2, ZrO2, TiO2, and Fe3O4, were used in thermodynamic analysis to derive the standard enthalpies of their surface protonation. Two different approaches were used for calculating the protonation enthalpy: one is based on thermodynamic description of the 1-pKa model for surface protonation, and another one on a combination of crystal chemistry and solvation theories which link the relative permittivity of the solid phase and the ratio of the Pauling bond strength and bond length to standard protonation enthalpy. From this analysis, two expressions relating the protonation enthalpy to the relative permittivity of the solid phase were obtained.

  3. Protonation enthalpies of metal oxides from high temperature electrophoresis.

    SciTech Connect

    Rodriguez-Santiago, V; Fedkin, Mark V; Lvov, Serguei N.

    2012-01-01

    Surface protonation reactions play an important role in the behavior of mineral and colloidal systems, specifically in hydrothermal aqueous environments. However, studies addressing the reactions at the solid/liquid interface at temperatures above 100 C are scarce. In this study, newly and previously obtained high temperature electrophoresis data (up to 260 C) - zeta potentials and isoelectric points - for metal oxides, including SiO{sub 2}, SnO{sub 2}, ZrO{sub 2}, TiO{sub 2}, and Fe{sub 3}O{sub 4}, were used in thermodynamic analysis to derive the standard enthalpies of their surface protonation. Two different approaches were used for calculating the protonation enthalpy: one is based on thermodynamic description of the 1-pKa model for surface protonation, and another one - on a combination of crystal chemistry and solvation theories which link the relative permittivity of the solid phase and the ratio of the Pauling bond strength and bond length to standard protonation enthalpy. From this analysis, two expressions relating the protonation enthalpy to the relative permittivity of the solid phase were obtained.

  4. Capillary electrophoresis of sialylated oligosaccharides in milk from different species.

    PubMed

    Monti, Lucia; Cattaneo, Tiziana Maria Piera; Orlandi, Mario; Curadi, Maria Claudia

    2015-08-28

    Oligosaccharides are relevant components of human milk, which have been quite well studied for their pre-biotic effect and their capacity in stimulating the immune system. Since oligosaccharides from milk of non-human mammals received so far less attention, the aim of this work was the application of capillary electrophoresis (CE) for the analysis of sialylated oligosaccharides in cow, goat and equine (mare and donkey) milk to possibly identify potential sources of oligosaccharides to use as health promoting ingredients in functional foods. Human milk was used as reference milk. A recent CE technique was applied to resolve and quantify 3-sialyllactose (3-SL), 6-sialyllactose (6-SL) and disialyl-lacto-N-tetraose (DSLNT). Analysis of non-human milk samples confirmed differences among species and individuals: DSLNT, which was the most abundant compound in human milk (455-805μg/mL) was missing in most of the samples. In most cases, 3-SL showed to be the most concentrated of the quantified analytes, with values ranging from 12 to 77μg/mL. PMID:26228851

  5. A Contactless Capacitance Detection System for Microchip Capillary Electrophoresis

    NASA Astrophysics Data System (ADS)

    Wu, Peter

    2008-05-01

    The design, construction and operation of a simple, inexpensive and compact high voltage power supply for use in conjunction with a simple cross, capillary electrophoresis microchip is presented. The detection system utilizes a single high voltage power supply (15 kV), a voltage divider network for obtaining the required voltages for enabling a gated injection valve, and two high voltage relays for switching between the open and closed gate sequences of the injection. The system is used to determine sodium monofluoroacetate (MFA) concentration in diluted fruit juices and tap water. A separation buffer consisting of 20 mM citric acid and histidine at pH 3.5 enabled the detection of the anion in diluted apple juice, cranberry juice, and orange juice without lengthy sample pretreatments. Limit of detection in diluted juices and tap water were determined to be 125, 167, 138, and 173 mg/L for tap water, apple juice, cranberry juice, and orange juice, respectively, based upon an S/N of 3:1. The total analysis time for detecting the MFA anion in fruit juices was less than 5 min, which represents a considerable reduction in analysis time compared to other analytical methods currently used in food analysis.

  6. Implementation of microchip electrophoresis instrumentation for future spaceflight missions.

    PubMed

    Willis, Peter A; Creamer, Jessica S; Mora, Maria F

    2015-09-01

    We present a comprehensive discussion of the role that microchip electrophoresis (ME) instrumentation could play in future NASA missions of exploration, as well as the current barriers that must be overcome to make this type of chemical investigation possible. We describe how ME would be able to fill fundamental gaps in our knowledge of the potential for past, present, or future life beyond Earth. Despite the great promise of ME for ultrasensitive portable chemical analysis, to date, it has never been used on a robotic mission of exploration to another world. We provide a current snapshot of the technology readiness level (TRL) of ME instrumentation, where the TRL is the NASA systems engineering metric used to evaluate the maturity of technology, and its fitness for implementation on missions. We explain how the NASA flight implementation process would apply specifically to ME instrumentation, and outline the scientific and technology development issues that must be addressed for ME analyses to be performed successfully on another world. We also outline research demonstrations that could be accomplished by independent researchers to help advance the TRL of ME instrumentation for future exploration missions. The overall approach described here for system development could be readily applied to a wide range of other instrumentation development efforts having broad societal and commercial impact. PMID:26253225

  7. Capillary electrophoresis of conidia from cultivated microscopic filamentous fungi.

    PubMed

    Horká, Marie; Růzicka, Filip; Kubesová, Anna; Holá, Veronika; Slais, Karel

    2009-05-15

    In immunocompromised people fungal agents are able to cause serious infections with high mortality rate. An early diagnosis can increase the chances of survival of the affected patients. Simultaneously, the fungi produce toxins and they are frequent cause of allergy. Currently, various methods are used for detection and identification of these pathogens. They use microscopic examination and growth characteristic of the fungi. New methods are based on the analysis of structural elements of the target microorganisms such as proteins, polysaccharides, glycoproteins, nucleic acids, etc. for the construction of antibodies, probes, and primers for detection. The above-mentioned methods are time-consuming and elaborate. Here hydrophobic conidia from the cultures of different strains of the filamentous fungi were focused and separated by capillary zone electrophoresis and capillary isoelectric focusing. The detection was optimized by dynamic modifying of conidia by the nonionogenic tenside on the basis of pyrenebutanoate. Down to 10 labeled conidia of the fungal strains were fluorometrically detected, and isoelectric points of conidia were determined. The observed isoelectric points were compared with those obtained from the separation of the cultured clinical samples, and they were found to be not host-specific. PMID:19385663

  8. Misincorporation during DNA synthesis, analyzed by gel electrophoresis.

    PubMed Central

    Hillebrand, G G; McCluskey, A H; Abbott, K A; Revich, G G; Beattie, K L

    1984-01-01

    A method has been developed for simultaneous comparison of the propensity of a DNA polymerase to misincorporate at different points on a natural template-primer. In this method elongation of a [5'-32P] primer, annealed to a bacteriophage template strand, is carried out in the presence of only three dNTPs (highly purified by HPLC). Under these conditions the rate of primer elongation (monitored by gel electrophoresis/autoradiography) is limited by the rate of misincorporation at template positions complementary to the missing dNTP. Variations in the rate of elongation (revealed by autoradiographic banding patterns) reflect variations in the propensity for misincorporation at different positions along the template. The effect on primer elongation produced by addition of a chemically modified dNTP to 'minus' reactions reveals the mispairing potential of the modified nucleotide during DNA synthesis. By use of this electrophoretic assay of misincorporation we have demonstrated that the fidelity of E. coli DNA polymerase I varies greatly at different positions along a natural template, and that BrdUTP and IodUTP can be incorporated in place of dCTP during chain elongation catalyzed by this enzyme. Images PMID:6326053

  9. Capillary electrophoresis microchip detecting system based on embedded optical fiber

    NASA Astrophysics Data System (ADS)

    Yan, Weiping; Li, Yuanyuan; Ma, Lingzhi

    2007-12-01

    Microchip capillary electrophoresis(CE) has been recognized as a powerful tool for biochemical analyses due to its smaller size, faster separation and lower sample requirement. According to the principle of laser-induced fluorescence, the detecting system of CE microchip embedded optical fiber is discussed in this paper as well as its small volume and simple detection optical circuit. The system was composed with semiconductor laser (532nm), high voltage control system, photon counter, PC and CE chip embedded optical fibers. With the constructed detection system, different samples and different concentrations were detected, including Rhodamine B, Rhodamine 6G, and mingling solution of Rhodamine B and Rhodamine 6G. The lowest detected concentration is 1×10 -6mol/L for Rhodamine B, and 1×10 -5mol/L for Rhodamine 6G, respectively. The separation of the mingling solution of Rhodamine B and Rhodamine 6G was completed, whose concentration were both about 1×10 -4mol/L. The results show that the constructed detection system possesses some advantages, such as compact structure, higher sensitivity and repetition, which are beneficial to the development of microminiaturization and integration of micro CE chip.

  10. Capillary Electrophoresis-Based Assay of Phosphofructokinase-1

    PubMed Central

    Malina, Andrew; Bryant, Sherrisse K.; Chang, Simon H.; Waldrop, Grover L.; Gilman, S. Douglass

    2013-01-01

    An assay was developed for phosphofructokinase-1 (PFK-1) using capillary electrophoresis (CE). In the glycolytic pathway, this enzyme catalyzes the rate-limiting step from fructose-6-phosphate and magnesium-bound adenosine triphosphate (Mg-ATP) to fructose-1,6-bisphosphate and magnesium-bound adenosine diphosphate (Mg-ADP). This enzyme has recently become a research target because of the importance of glycolysis in cancer and obesity. The CE assay for PFK-1 is based on the separation and detection by UV absorbance at 260 nm of Mg-ATP and Mg-ADP. The separation was enhanced by addition of Mg2+ to the separation buffer. Inhibition studies of PFK-1 by aurintricarboxylic acid and palmitoyl coenzyme A were also performed. An IC50 value was determined for aurintricarboxylic acid, and this value matched values in the literature obtained using coupled spectrophotometric assays. This assay for PFK-1 directly monitors the enzyme-catalyzed reaction, and the CE separation reduces the potential of spectral interference by inhibitors. PMID:24444856

  11. Latex samples for RAMSES electrophoresis experiment on IML 2

    NASA Technical Reports Server (NTRS)

    Seaman, Geoffrey V. F.; Knox, Robert J.

    1994-01-01

    The objectives of these reported studies were to provide ground based support services for the flight experiment team for the RAMSES experiment to be flown aboard IML-2. The specific areas of support included consultation on the performance of particle based electrophoresis studies, development of methods for the preparation of suitable samples for the flight hardware, the screening of particles to obtain suitable candidates for the flight experiment, and the electrophoretic characterization of sample particle preparations. The first phases of these studies were performed under this contract, while the follow on work was performed under grant number NAG8 1081, 'Preparation and Characterization of Latex Samples for RAMSES Experiment on IML 2.' During this first phase of the experiment the following benchmarks were achieved: Methods were tested for the concentration and resuspension of latex samples in the greater than 0.4 micron diameter range to provide moderately high solids content samples free of particle aggregation which interferred with the normal functioning of the RAMSES hardware. Various candidate latex preparations were screened and two candidate types of latex were identified for use in the flight experiments, carboxylate modified latex (CML) and acrylic acid-acrylamide modified latex (AAM). These latexes have relatively hydrophilic surfaces, are not prone to aggregate, and display sufficiently low electrophoretic mobilities in the flight buffer so that they can be used to make mixtures to test the resolving power of the flight hardware.

  12. DNA Length Ranges Exhibiting Distinct Separation Mechanisms in Gel Electrophoresis

    NASA Astrophysics Data System (ADS)

    Beheshti, A.; van Winkle, D. H.; Rill, R. L.

    2003-03-01

    Electrophoresis was performed on double stranded DNA ranging from 200 to 194,000 bp in agarose gel concentrations from 0.4% - 1.3%. The electric field was varied from 0.62 to 6.21 V/cm. A wide range of electric fields and gel concentrations were used to study how the new interpolation equation, frac1μ(L) = frac1μL - (frac1μL - frac1μ_s)e^-L/γ (where μ_L, μ_s, and γ are independent free fitting parameters), helps to distinguish among different mechanisms of molecular transport. This exponential relation fits well when there is a smooth transition from Ogston sieving to reptation. These transitions are distinguished by so-called ``reptation plots" (plotting 3μ L/μ_rc vs. L) (J. Rousseau, G. Drouin, and G. W. Slater, Phys Rev Lett. 1997, 79, 1945-1948). Fits deviate from the data more than two characteristic trends are observed in the reptation plots. The failure of the fits to follow the data appears to be a consequence of another separation mechanism, ``entropic trapping," occurring between the sieving and reptation regimes. The boundaries between length and field ranges where different separation mechanisms dominate are extracted from reptation plots of the best fits and the data. ``Phase diagrams" expressing these boundaries are derived.

  13. Spot identification on 2D electrophoresis gel images

    NASA Astrophysics Data System (ADS)

    Wang, Weixing

    2006-09-01

    2-D electrophoresis gel images can be used for identifying and characterizing many forms of a particular protein encoded by a single gene. Conventional approaches to gel analysis require the three steps: (1) Spot detection on each gel; (2) Spot matching between gels; and (3) Spot quantification and comparison. Many researchers and developers attempt to automate all steps as much as possible, but errors in the detection and matching stages are common. In order to carry out gel image analysis, one first needs to accurately detect and measure the protein spots in a gel image. This paper presents the algorithms for automatically delineating gel spots. The fusion of two types of segmentation algorithms was implemented. One is edge (discontinuity) based type, and the other is region based type. The primary integration of the two types of image segmentation algorithms have been tested too, the test results clearly show that the integrated algorithm can automatically delineate gel spots not only on a simple image and also on a complex image, and it is much better that either only edge based algorithm or only region based algorithm. Based on the testing and analysis results, the fusion of edge information and region information for gel image segmentation is good for this kind of images.

  14. DNA walks one step at a time in electrophoresis

    NASA Astrophysics Data System (ADS)

    Guan, Juan; Wang, Bo; Granick, Steve

    2011-03-01

    Testing the classical view that in DNA gel electrophoresis, long polymer chains navigate through their gel environment via reptation, we reach a different conclusion: this driven motion proceeds by stick-slip. Our single-molecule experiments visualize fluorescent-labeled lambda-DNA, whose intramolecular conformations are resolved with 30 ms resolution using home-written software. Combining hundreds to thousands of trajectories under amplitudes of electric field ranging from zero to large, we quantify the full statistical distribution of motion with unprecedented statistics. Pauses are seen between steps of driven motion, probably reflecting that the chain is trapped inside the gel matrix. The pausing time is exponentially distributed and decreases with increasing electric field strength, suggesting that the jerky behavior is an activated process, facilitated by electric field. We propose a stretch-assisted mechanism: that the energy barrier to move through the gel environment is first overcome by a leading segment, the ensuing intramolecular stress from stretching causing lagging segments to recoil and follow along.

  15. A vertical submarine electrophoresis apparatus for polyacrylamide minigels.

    PubMed

    Spiker, S

    1990-03-01

    A vertical submarine electrophoresis apparatus for use with minislab polyacrylamide gels is described. The design allows polyacrylamide gels to be run with the same ease and convenience that agarose gels are run with horizontal submarine apparatuses. The vertical submarine features a single buffer chamber with a restriction between the upper and the lower portions of the chamber. Acrylamide gels, cast between 9 X 10-cm glass slides, are inserted into the restriction and are completely immersed in buffer. Thus, current flows primarily through the gel itself, but some current flows through the buffer in the restriction surrounding the gel. Because water-tight separation of buffer chambers is not necessary, time-consuming and/or expensive procedures such as sealing with agarose or using fragile notched glass plates are eliminated. The apparatus can be set up to run a gel in less than 30 s. It is versatile in that gels of varying thickness (0.5, 0.8, 1.5, and 3 mm) can be run on a single apparatus. The apparatus has been used for sodium dodecyl sulfate gels, low ionic strength native gels for nucleoprotein complexes, and composite acrylamide-agarose gels. PMID:2339782

  16. Analysis of results of ASTP experiment in electrophoresis

    NASA Technical Reports Server (NTRS)

    Vanderhoff, J. W.; Micale, F. J.; Krumrine, P. H.

    1977-01-01

    The Apollo-Soyuz Test Project (ASTP) included an electrophoretic separation experiment of biological cells. The nature separation results of aldehyde-fixed rabbit, human and horse red blood cells, which were taken in the form of photographs taken at three-minute intervals, are the subject of this report. The electrophoretic separation was successful in that fractionation according to mobility did occur and was found in the sliced samples. Photographic evidence indicates that the low electroosmotic methylcellulose coating was successful in reducing the electroosmosis to a near zero value. Also, the flight film shows that the bands migrated down the column as theory would predict, producing two bands of high cell concentration separated and surrounded by regions of lower cell concentration. However, most likely some clumping of cells occurred to cause the trailing band to be larger than expected from theory. Overall, the experiment was a success in demonstrating a static electrophoresis separation under microgravity conditions with a resolution not possible on earth.

  17. Diagnostics of Titan's stratospheric dynamics using Cassini/CIRS data and the 2-dimensional IPSL circulation model

    NASA Astrophysics Data System (ADS)

    Crespin, A.; Lebonnois, S.; Vinatier, S.; Bézard, B.; Coustenis, A.; Teanby, N. A.; Achterberg, R. K.; Rannou, P.; Hourdin, F.

    2008-10-01

    The dynamics of Titan's stratosphere is discussed in this study, based on a comparison between observations by the CIRS instrument on board the Cassini spacecraft, and results of the 2-dimensional circulation model developed at the Institute Pierre-Simon Laplace, available at http://www.lmd.jussieu.fr/titanDbase [Rannou, P., Lebonnois, S., Hourdin, F., Luz, D., 2005. Adv. Space Res. 36, 2194-2198]. The comparison aims at both evaluating the model's capabilities and interpreting the observations concerning: (1) dynamical and thermal structure using temperature retrievals from Cassini/CIRS and the vertical profile of zonal wind at the Huygens landing site obtained by Huygens/DWE; and (2) vertical and latitudinal profiles of stratospheric gases deduced from Cassini/CIRS data. The modeled thermal structure is similar to that inferred from observations (Cassini/CIRS and Earth-based observations). However, the upper stratosphere (above 0.05 mbar) is systematically too hot in the 2D-CM, and therefore the stratopause region is not well represented. This bias may be related to the haze structure and to misrepresented radiative effects in this region, such as the cooling effect of hydrogen cyanide (HCN). The 2D-CM produces a strong atmospheric superrotation, with zonal winds reaching 200 m s -1 at high winter latitudes between 200 and 300 km altitude (0.1-1 mbar). The modeled zonal winds are in good agreement with retrieved wind fields from occultation observations, Cassini/CIRS and Huygens/DWE. Changes to the thermal structure are coupled to changes in the meridional circulation and polar vortex extension, and therefore affect chemical distributions, especially in winter polar regions. When a higher altitude haze production source is used, the resulting modeled meridional circulation is weaker and the vertical and horizontal mixing due to the polar vortex is less extended in latitude. There is an overall good agreement between modeled chemical distributions and observations

  18. A large-scale electrophoresis- and chromatography-based determination of gene expression profiles in bovine brain capillary endothelial cells after the re-induction of blood-brain barrier properties

    PubMed Central

    2010-01-01

    Background Brain capillary endothelial cells (BCECs) form the physiological basis of the blood-brain barrier (BBB). The barrier function is (at least in part) due to well-known proteins such as transporters, tight junctions and metabolic barrier proteins (e.g. monoamine oxidase, gamma glutamyltranspeptidase and P-glycoprotein). Our previous 2-dimensional gel proteome analysis had identified a large number of proteins and revealed the major role of dynamic cytoskeletal remodelling in the differentiation of bovine BCECs. The aim of the present study was to elaborate a reference proteome of Triton X-100-soluble species from bovine BCECs cultured in the well-established in vitro BBB model developed in our laboratory. Results A total of 215 protein spots (corresponding to 130 distinct proteins) were identified by 2-dimensional gel electrophoresis, whereas over 350 proteins were identified by a shotgun approach. We classified around 430 distinct proteins expressed by bovine BCECs. Our large-scale gene expression analysis enabled the correction of mistakes referenced into protein databases (e.g. bovine vinculin) and constitutes valuable evidence for predictions based on genome annotation. Conclusions Elaboration of a reference proteome constitutes the first step in creating a gene expression database dedicated to capillary endothelial cells displaying BBB characteristics. It improves of our knowledge of the BBB and the key proteins in cell structures, cytoskeleton organization, metabolism, detoxification and drug resistance. Moreover, our results emphasize the need for both appropriate experimental design and correct interpretation of proteome datasets. PMID:21078152

  19. Step width, spacing, and resolution in gradient elution moving boundary electrophoresis. Part 1. Theory and comparison with zone electrophoresis.

    PubMed

    Ross, David

    2010-11-01

    Gradient elution moving boundary electrophoresis (GEMBE) is a recently described technique for electrophoretic separations in short capillaries or microchannels. With GEMBE, the electrophoretic migration of analytes is opposed by a bulk counterflow of separation buffer. The counterflow velocity is varied over the course of a separation so that analytes with different mobilities enter the separation channel at different times and are detected as moving boundary, step-wise increases in the detector response. The resolution of a GEMBE separation is thus dependent on the rate at which the counterflow velocity is varied, and relatively high-resolution separations can be performed with short microfluidic channels or capillaries. In this paper, a theoretical description of GEMBE is presented that can be used for calculation of expected resolution and for optimization of GEMBE separations. A comparison is made with CZE, a conventional electrophoretic separation technique for which the theoretical understanding is mature. The results indicate that the electric field strength and separation channel length are important parameters for both CZE and GEMBE. However, with GEMBE, the counterflow acceleration is also available as a parameter that can be easily adjusted for optimization of the trade-off between resolution and separation time. This allows for optimization of GEMBE separations by making changes only to the software controlling the apparatus rather than to the hardware of the apparatus itself. Further comparison of the theoretical descriptions of GEMBE and CZE indicates that the time required to achieve a desired resolution is equivalent for the two techniques. PMID:21077236

  20. Renaturation of enzymes after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate

    SciTech Connect

    Lacks, S.A.; Springhorn, S.S.

    1980-08-10

    A number of enzymes, including amylases, dehydrogenases, and proteases, were shown to be renaturable after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Enzyme activity was detected in situ by action on substrates introduced into the gel and subsequent staining of either the product or unreacted substrate. Enzymes appeared to recover activity as soon as the detergent diffused out of the gel. Renatured enzymes were retained in gels after electrophoresis longer than native enzymes which had been subjected to electrophoresis in the absence of detergent. Re-electrophoresis of the renatured enzymes showed that part of the retained activity was physically anchored to the gel, possibly by the folding of polypeptides around the gel matrix as the enzymes were renatured.

  1. Thiocyanato Chromium (III) Complexes: Separation by Paper Electrophoresis and Estimate of Stability Constants

    ERIC Educational Resources Information Center

    Larsen, Erik; Eriksen, J.

    1975-01-01

    Describes an experiment wherein the student can demonstrate the existence of all the thiocyanato chromium complexes, estimate the stepwise formation constants, demonstrate the robustness of chromium III complexes, and show the principles of paper electrophoresis. (GS)

  2. APPLICATION OF ELECTROPHORESIS TO STUDY THE ENANTIOSELECTIVE TRANSFORMATION OF FIVE CHIRAL PESTICIDES IN AEROBIC SOIL SLURRIES

    EPA Science Inventory

    The enantiomers of five chiral pesticides of environmental interest, metalaxyl, imazaquin, fonofos (dyfonate), ruelene (cruformate) and dichlorprop, were separated analytically using capillary electrophoresis (CE) with cyclodextrin chiral selectors. CE is shown to be a simple, ef...

  3. Enzymatic membranes for the selective transport of neutral molecules by electrophoresis.

    PubMed

    Perrin, Bernard; Couturier, Roger; Fiaty, Koffi; Charcosset, Catherine; Maïsterrena, Bernard

    2008-06-01

    The active and selective transport of glucose and glycerol was carried out using electrophoresis and artificial enzymatic membranes. These positively charged composite membranes carry, on the face adjacent to the donor compartment of an electrophoresis module, a specific kinase (hexokinase or glycerokinase) and, on the opposite face, an alkaline phosphatase (ALP). Phosphorylation of the neutral substrate (glucose or glycerol) on the donor side by the kinase generates a negatively charged phosphorylated substrate, whose transmembrane migration is promoted by an electric field and by the membrane's positive charge. Dephosphorylation of the phosphorylated substrate by ALP on the opposite face regenerates the neutral substrate, which accumulates in the receiver compartment of the electrophoresis module. Using an electrophoresis module specifically designed for this study, our experiments were carried out enabling glucose and glycerol to be concentrated approximately eight- and twelve-fold, respectively, in 8 h. PMID:18435500

  4. APPLICATIONS OF CAPILLARY ELECTROPHORESIS/LASER-INDUCED FLUORESCENCE DETECTION TO GROUND WATER MIGRATION STUDIES

    EPA Science Inventory

    Capillary electrophoresis (CE) has been applied to the determination of groundwater migration based on laser-induced fluorescence (LIF) detection and traditional spectrofluorimetry. The detection limits of injected dye-fluorescent whitening agent (tinopal) in the low parts per tr...

  5. Capillary Electrophoresis Profiles and Fluorophore Components of Humic Acids in Nebraska Corn and Philippine Rice Soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As humic substances represent relatively high molecular mass polyelectrolytes containing aromatic, aliphatic and heterocyclic subunits, capillary electrophoresis (CE) has become an attractive method for “finger-print” characterization of humic acids. In addition, fluorescence excitation-emission ma...

  6. ANALYSIS OF THE ENANTIOMERS OF CHIRAL PESTICIDES AND OTHER POLLUTANTS IN ENVIRONMENTAL SAMPLES BY CAPILLARY ELECTROPHORESIS

    EPA Science Inventory

    The generic method described here involves typical capillary electrophoresis (CE) techniques, with the addition of cyclodextrin chiral selectors to the electrolyte for enantiomer separation and also, in the case of neutral analytes, the further addition of a micelle forming comp...

  7. Electrophoresis tests on STS-3 and ground control experiments - A basis for future biological sample selections

    NASA Technical Reports Server (NTRS)

    Morrison, D. R.; Lewis, M. L.

    1982-01-01

    Static zone electrophoresis is an electrokinetic method of separating macromolecules and small particles. However, its application for the isolation of biological cells and concentrated protein solutions is limited by sedimentation and convection. Microgravity eliminates or reduces sedimentation, floatation, and density-driven convection arising from either Joule heating or concentration differences. The advantages of such an environment were first demonstrated in space during the Apollo 14 and 16 missions. In 1975 the Electrophoresis Technology Experiment (MA-011) was conducted during the Apollo-Soyuz Test Project flight. In 1979 a project was initiated to repeat the separations of human kidney cells. One of the major objectives of the Electrophoresis Equipment Verification Tests (EEVT) on STS-3 was to repeat and thereby validate the first successful electrophoretic separation of human kidney cells. Attention is given to the EEVT apparatus, the preflight electrophoresis, and inflight operational results.

  8. IDENTIFICATION OF REACTIVE DYES IN SPENT DYEBATHS AND WASTEWATER BY CAPILLARY ELECTROPHORESIS/MASS SPECTROMETRY

    EPA Science Inventory

    Capillary electrophoresis with diode array detection and mass spectrometry combined with solid-phase extraction were employed for the identification of reactive vinylsulfone and chlorotriazine dyes and their hydrolysis products in spent dyebaths and raw and treated wastewater. Re...

  9. INFLUENCE OF BORATE BUFFERS ON THE ELECTROPHORETIC BEHAVIOR OF HUMIC SUBSTANCES IN CAPILLARY ZONE ELECTROPHORESIS

    EPA Science Inventory

    The influence of tetrahydroxyborate ions on the electrophoretic mobility of humic acids was evaluated by capillary electrophoresis (CE). Depending on the molarity of borate ions in the separation buffer, the humic acids exhibit electropherograms with sharp peaks consistently exte...

  10. Estimation of circular DNA size using gamma-irradiation and pulsed-field gel electrophoresis

    SciTech Connect

    Beverley, S.M. )

    1989-02-15

    A method is described for estimating the size of large circular DNAs found within complex chromosomal DNA preparations. DNAs are treated with low levels of gamma-irradiation, sufficient to introduce a single double-stranded break per circle, and the resulting linear DNA is sized by pulsed-field electrophoresis and blot hybridization. The method is fast, reproducible, and very conveniently applied to the agarose-enclosed chromosomal DNA preparations commonly used in pulsed field electrophoresis.

  11. [The effect of magnesium sulfate electrophoresis and galvanization on the mineralization of teeth and bones].

    PubMed

    Varava, G N; Podorozhnaia, R P; Genesina, T I; Sukmanskiĭ, V B

    1990-01-01

    Effects of Mg2+ electrophoresis and galvanization on tooth and bone mineralization was experimentally studied with the use of radioactive Ca and P isotopes. Mg2+ electrophoresis and, to a lesser degree, galvanization enhanced 32P incorporation in incisors and maxillary bones. Mg2+ significantly increased 45Ca incorporation in teeth and maxillary bones. Experimental data permit clinical trials of Mg2+ efficacy in patients with disordered mineralization and remineralization. PMID:2389264

  12. Chemiluminescence resonance energy transfer-based detection for microchip electrophoresis.

    PubMed

    Zhao, Shulin; Huang, Yong; Shi, Ming; Liu, Rongjun; Liu, Yi-Ming

    2010-03-01

    Since the channels in micro- and nanofluidic devices are extremely small, a sensitive detection is required following microchip electrophoresis (MCE). This work describes a highly sensitive and yet universal detection scheme based on chemiluminescence resonance energy transfer (CRET) for MCE. It was found that an efficient CRET occurred between a luminol donor and a CdTe quantum dot (QD) acceptor in the luminol-NaBrO-QD system and that it was sensitively suppressed by the presence of certain organic compounds of biological interest including biogenic amines and thiols, amino acids, organic acids, and steroids. These findings allowed developing sensitive MCE-CL assays for the tested compounds. The proposed MCE-CL methods showed desired analytical figures of merit such as a wide concentration range of linear response. Detection limits obtained were approximately 10(-9) M for biogenic amines including dopamine and epinephrine and approximately 10(-8) M for biogenic thiols (e.g., glutathione and acetylcysteine), organic acids (i.e., ascorbic acid and uric acid), estrogens, and native amino acids. These were 10-1000 times more sensitive than those of previously reported MCE-based methods with chemiluminescence, electrochemical, or laser-induced fluorescence detection for quantifying corresponding compounds. To evaluate the applicability of the present MCE-CL method for analyzing real biological samples, it was used to determine amino acids in individual human red blood cells. Nine amino acids, including Lys, Ser, Ala, Glu, Trp, etc., were detected. The contents ranged from 3 to 31 amol/cell. The assay proved to be simple, quick, reproducible, and very sensitive. PMID:20121202

  13. Insight of Saffron Proteome by Gel-Electrophoresis.

    PubMed

    Paredi, Gianluca; Raboni, Samanta; Marchesani, Francesco; Ordoudi, Stella A; Tsimidou, Maria Z; Mozzarelli, Andrea

    2016-01-01

    Saffron is a spice comprised of the dried stigmas and styles of Crocus sativus L. flowers and, since it is very expensive, it is frequently adulterated. So far, proteomic tools have never been applied to characterize the proteome of saffron or identify possible cases of fraud. In this study, 1D-Gel Electrophoresis was carried out to characterize the protein profile of (i) fresh stigmas and styles of the plant; (ii) dried stigmas and styles from different geographical origins (Spanish, Italian, Greek and Iranian) that had been stored for various periods of time after their processing; and (iii) two common plant adulterants, dried petals of Carthamus tinctorius L. and dried fruits of Gardenia jasminoides Ellis. A selective protein extraction protocol was applied to avoid interference from colored saffron metabolites, such as crocins, during electrophoretic analyses of saffron. We succeeded in separating and assigning the molecular weights to more than 20 proteins. In spite of the unavailability of the genome of saffron, we were able to identify five proteins by Peptide Mass Fingerprinting: phosphoenolpyruvate carboxylase 3, heat shock cognate 70 KDa protein, crocetin glucosyltransferase 2, α-1,4-glucan-protein synthase and glyceraldehydes-3-phosphate dehydrogenase-2. Our findings indicate that (i) few bands are present in all saffron samples independently of origin and storage time, with amounts that significantly vary among samples and (ii) aging during saffron storage is associated with a reduction in the number of detectable bands, suggesting that proteases are still active. The protein pattern of saffron was quite distinct from those of two common adulterants, such as the dried petals of Carthamus tinctorius and the dried fruits of Gardenia jasminoides indicating that proteomic analyses could be exploited for detecting possible frauds. PMID:26840283

  14. Rapid inorganic ion analysis using quantitative microchip capillary electrophoresis.

    PubMed

    Vrouwe, Elwin X; Luttge, Regina; Olthuis, Wouter; van den Berg, Albert

    2006-01-13

    Rapid quantitative microchip capillary electrophoresis (CE) for online monitoring of drinking water enabling inorganic ion separation in less than 15 s is presented. Comparing cationic and anionic standards at different concentrations the analysis of cationic species resulted in non-linear calibration curves. We interpret this effect as a variation in the volume of the injected sample plug caused by changes of the electroosmotic flow (EOF) due to the strong interaction of bivalent cations with the glass surface. This explanation is supported by the observation of severe peak tailing. Conducting microchip CE analysis in a glass microchannel, optimized conditions are received for the cationic species K+, Na+, Ca2+, Mg2+ using a background electrolyte consisting of 30 mmol/L histidine and 2-(N-morpholino)ethanesulfonic acid, containing 0.5 mmol/L potassium chloride to reduce surface interaction and 4 mmol/L tartaric acid as a complexing agent resulting in a pH-value of 5.8. Applying reversed EOF co-migration for the anionic species Cl-, SO42- and HCO3- optimized separation occurs in a background electrolyte consisting of 10 mmol/L 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) and 10 mmol/L HEPES sodium salt, containing 0.05 mmol/L CTAB (cetyltrimethylammonium bromide) resulting in a pH-value of 7.5. The detection limits are 20 micromol/L for the monovalent cationic and anionic species and 10 micromol/L for the divalent species. These values make the method very suitable for many applications including the analysis of abundant ions in tap water as demonstrated in this paper. PMID:16310794

  15. Weakly nonlinear electrophoresis of a highly charged colloidal particle

    NASA Astrophysics Data System (ADS)

    Schnitzer, Ory; Zeyde, Roman; Yavneh, Irad; Yariv, Ehud

    2013-05-01

    At large zeta potentials, surface conduction becomes appreciable in thin-double-layer electrokinetic transport. In the linear weak-field regime, where this effect is quantified by the Dukhin number, it is manifested in non-Smoluchowski electrophoretic mobilities. In this paper we go beyond linear response, employing the recently derived macroscale model of Schnitzer and Yariv ["Macroscale description of electrokinetic flows at large zeta potentials: Nonlinear surface conduction," Phys. Rev. E 86, 021503 (2012), 10.1103/PhysRevE.86.021503] as the infrastructure for a weakly nonlinear analysis of spherical-particle electrophoresis. A straightforward perturbation in the field strength is frustrated by the failure to satisfy the far-field conditions, representing a non-uniformity of the weak-field approximation at large distances away from the particle, where salt advection becomes comparable to diffusion. This is remedied using inner-outer asymptotic expansions in the spirit of Acrivos and Taylor ["Heat and mass transfer from single spheres in Stokes flow," Phys. Fluids 5, 387 (1962), 10.1063/1.1706630], with the inner region representing the particle neighborhood and the outer region corresponding to distances scaling inversely with the field magnitude. This singular scheme furnishes an asymptotic correction to the electrophoretic velocity, proportional to the applied field cubed, which embodies a host of nonlinear mechanisms unfamiliar from linear electrokinetic theories. These include the effect of induced zeta-potential inhomogeneity, animated by concentration polarization, on electro-osmosis and diffuso-osmosis; bulk advection of salt; nonuniform bulk conductivity; Coulomb body forces acting on bulk volumetric charge; and the nonzero electrostatic force exerted upon the otherwise screened particle-layer system. A numerical solution of the macroscale model validates our weakly nonlinear analysis.

  16. A versatile polyacrylamide gel electrophoresis based sulfotransferase assay

    PubMed Central

    2010-01-01

    Background Sulfotransferases are a large group of enzymes that regulate the biological activity or availability of a wide spectrum of substrates through sulfation with the sulfur donor 3'-phosphoadenosine-5'-phosphosulfate (PAPS). These enzymes are known to be difficult to assay. A convenient assay is needed in order to better understand these enzymes. Results A universal sulfotransferase assay method based on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is described. This assay has been successfully applied to substrates as small as α-naphthol and as big as proteoglycans. As examples, we present the assays for recombinant human CHST4, TPST1, CHST3 and HS6ST1. In order to assess whether a small molecule can be applicable to this type of assay, a method to estimate the relative mobility of a molecule to PAPS is also presented. The estimated relative mobilities of various sulfated small molecules generated by SULT1A1, SULT1E1, SULT2A1 and CHST4 are in the range of ± 0.2 of the actual relative mobilities. Conclusion The versatility of the current method comes from the ability that SDS-PAGE can separate proteins and small molecules according to different parameters. While mobilities of proteins during SDS-PAGE are inversely related to their sizes, mobilities of small molecules are positively related to their charge/mass ratios. The predicted relative mobility of a product to PAPS is a good indicator of whether a sulfotransferase can be assayed with SDS-PAGE. Because phosphorylation is most similar to sulfation in chemistry, the method is likely to be applicable to kinases as well. PMID:20146816

  17. A stability indicating capillary electrophoresis method for analysis of buserelin.

    PubMed

    Tamizi, Elnaz; Kenndler, Ernst; Jouyban, Abolghasem

    2014-01-01

    A simple and rapid stability indicating method based on capillary zone electrophoresis has been developed and validated for the analysis of buserelin (BUS). The best separations were achieved by using a bare fused silica capillary (75 μm i.d.; 65.5 cm total, 57.0 cm effective length), phosphate buffer (pH = 3.00; 26.4 mM), at 35 °C. The sample was hydrodynamically injected at 50 mbar for 5 seconds; the applied voltage was 30 kV and detection was carried out by UV-absorbance at 200 nm. Method validation resulted in the following figures of merit : the method was linear in the concentration range between 0.781 and 500 μg/mL (linear regression coefficient 0.9996), accuracy was between 99.3% and 100.9%, intra assay precision was between 0.3% and 1.0% and intermediate precision was between 1.0% and 2.1%. Evaluation of the specificity of the method showed no interference between excipients or products of force degradation and BUS. Under the selected conditions, separation of BUS and its degradation products was completed in less than 10 min, and BUS could be quantified after different stress conditions without any interference. The results enabled the conclusion that under thermal stress upon exposure to 90 °C BUS is degraded by first order kinetics. It was demonstrated that the method can be applied as a rapid and easy to use method for quantification and stability testing of BUS in biopharmaceutical formulations in quality control laboratories. PMID:25276180

  18. 3D Printed Micro Free-Flow Electrophoresis Device.

    PubMed

    Anciaux, Sarah K; Geiger, Matthew; Bowser, Michael T

    2016-08-01

    The cost, time, and restrictions on creative flexibility associated with current fabrication methods present significant challenges in the development and application of microfluidic devices. Additive manufacturing, also referred to as three-dimensional (3D) printing, provides many advantages over existing methods. With 3D printing, devices can be made in a cost-effective manner with the ability to rapidly prototype new designs. We have fabricated a micro free-flow electrophoresis (μFFE) device using a low-cost, consumer-grade 3D printer. Test prints were performed to determine the minimum feature sizes that could be reproducibly produced using 3D printing fabrication. Microfluidic ridges could be fabricated with dimensions as small as 20 μm high × 640 μm wide. Minimum valley dimensions were 30 μm wide × 130 μm wide. An acetone vapor bath was used to smooth acrylonitrile-butadiene-styrene (ABS) surfaces and facilitate bonding of fully enclosed channels. The surfaces of the 3D-printed features were profiled and compared to a similar device fabricated in a glass substrate. Stable stream profiles were obtained in a 3D-printed μFFE device. Separations of fluorescent dyes in the 3D-printed device and its glass counterpart were comparable. A μFFE separation of myoglobin and cytochrome c was also demonstrated on a 3D-printed device. Limits of detection for rhodamine 110 were determined to be 2 and 0.3 nM for the 3D-printed and glass devices, respectively. PMID:27377354

  19. Quantification of Carbohydrates in Grape Tissues Using Capillary Zone Electrophoresis

    PubMed Central

    Zhao, Lu; Chanon, Ann M.; Chattopadhyay, Nabanita; Dami, Imed E.; Blakeslee, Joshua J.

    2016-01-01

    Soluble sugars play an important role in freezing tolerance in both herbaceous and woody plants, functioning in both the reduction of freezing-induced dehydration and the cryoprotection of cellular constituents. The quantification of soluble sugars in plant tissues is, therefore, essential in understanding freezing tolerance. While a number of analytical techniques and methods have been used to quantify sugars, most of these are expensive and time-consuming due to complex sample preparation procedures which require the derivatization of the carbohydrates being analyzed. Analysis of soluble sugars using capillary zone electrophoresis (CZE) under alkaline conditions with direct UV detection has previously been used to quantify simple sugars in fruit juices. However, it was unclear whether CZE-based methods could be successfully used to quantify the broader range of sugars present in complex plant extracts. Here, we present the development of an optimized CZE method capable of separating and quantifying mono-, di-, and tri-saccharides isolated from plant tissues. This optimized CZE method employs a column electrolyte buffer containing 130 mM NaOH, pH 13.0, creating a current of 185 μA when a separation voltage of 10 kV is employed. The optimized CZE method provides limits-of-detection (an average of 1.5 ng/μL) for individual carbohydrates comparable or superior to those obtained using gas chromatography–mass spectrometry, and allows resolution of non-structural sugars and cell wall components (structural sugars). The optimized CZE method was successfully used to quantify sugars from grape leaves and buds, and is a robust tool for the quantification of plant sugars found in vegetative and woody tissues. The increased analytical efficiency of this CZE method makes it ideal for use in high-throughput metabolomics studies designed to quantify plant sugars. PMID:27379118

  20. Thermodynamics and ideal glass transition on the surface of a monatomic system modeled as quasi "2-dimensional" recursive lattices

    NASA Astrophysics Data System (ADS)

    Huang, Ran

    Two quasi 2-dimensional recursive lattices formed by planar elements have been designed to investigate the surface thermodynamics of monatomic Ising glass system with the aim to study the metastability of supercooled liquids and the ideal glass transition. Both lattices are constructed as hybrids of a Husimi lattice representing the bulk and lower dimensional recursive trees representing the surface. The coordination number, i.e. the number of neighbor sites surrounding one site, is designed to be 3 on the surface and 4 inside the bulk to mimic the 2D regular square lattice case. The recursive properties of recursive lattices were adopted to obtain exact thermodynamic calculations without approximation. The model has a strong anti-ferromagnetic interaction to give rise to an ordered phase identified as a crystal, and a metastable solution is also found to represent the amorphous phase. Interactions between particles farther away than the nearest neighbor distance are taken into consideration. The calculations were done with C/C++ programs. A recursive calculation technique was employed to approach an exact description of the system with the ratio of partial partition functions (PPF) on each site of the lattice. Thermal properties including free energy, energy density and entropy of the ideal crystal and supercooled liquid state of the model on the surface are calculated by the PPF. By analyzing the free energies and entropies of the crystal and supercooled liquid state, we are able to identify the melting transition and the second order ideal glass transition on the surface. The effects of different energy terms that produce competitions between crystallization and glass transition are studied. The results show that due to the coordination number change, the transition temperature on the surface decreases significantly compared to the transition temperature of the bulk system obtained in our previous research. Our theoretical calculation agrees with experiments and

  1. Modification of vital wheat gluten with phosphoric acid to produce high free-solution capacity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wheat gluten reacts with phosphoric acid to produce natural superabsorbent gels. The gel properties are defined by Fourier Transform Infra-red (FTIR) spectroscopy, 2-dimensional gel electrophoresis (2DE), and uptake of water, salt solutions, and aqueous ethanol. Temperatures above 120'C and dry cond...

  2. ISOFORM-SPECIFIC BINDING OF 14-3-3 PROTEINS TO NITRATE REDUCTASE AND THE BRASSINOSTEROID INSENSITIVE 1 RECEPTOR KINASE SIGNALING COMPLEX

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 14-3-3 proteins are known to bind many different soluble protein clients, but less is known about binding to integral membrane proteins, and in both cases the issue of isoform specificity remains largely unexplored. Using an array of anti-14-3-3 antibodies and 2-dimensional electrophoresis (2-DE...

  3. Separation of fluorescently labeled phosphoinositides and sphingolipids by capillary electrophoresis

    PubMed Central

    Wang, Kelong; Jiang, Dechen; Sims, Christopher E.; Allbritton, Nancy L.

    2012-01-01

    Phosphoinositides (PIs) and sphingolipids regulate many aspects of cell behavior and are often involved in disease processes such as oncogenesis. Capillary electrophoresis with laser induced fluorescence detection (CE-LIF) is emerging as an important tool for enzymatic assays of the metabolism of these lipids, particularly in cell-based formats. Previous separations of phosphoinositide lipids by CE required a complex buffer with polymer additives which had the disadvantages of high cost and/or short shelf life. Further a simultaneous separation of these classes of lipids has not been demonstrated in a robust buffer system. In the current work, a simple separation buffer based on NaH2PO4 and 1-propanol was optimized to separate two sphingolipids and multiple phosphoinositides by CE. The NaH2PO4 concentration, pH, 1-propanol fraction, and a surfactant additive to the buffer were individually optimized to achieve simultaneous separation of the sphingolipids and phosphoinositides. Fluorescein-labeled sphingosine (SFL) and sphingosine 1-phosphate (S1PFL), fluorescein-labeled phosphatidyl-inositol 4,5-bisphosphate (PIP2) and phosphatidyl-inositol 3,4,5-trisphosphate (PIP3), and bodipy-fluorescein (BFL)-labeled PIP2 and PIP3 were separated pairwise and in combination to demonstrate the generalizability of the method. Theoretical plate numbers achieved were as high as 2×105 in separating fluorophore-labeled PIP2 and PIP3. Detection limits for the 6 analytes were in the range of 10−18 to 10−20 mol. The method also showed high reproducibility, as the relative standard deviation of the normalized migration time for each analyte in the simultaneous separation of all 6 compounds was less than 1%. The separation of a mixture composed of diacylglycerol (DAG) and multiple phosphoinositides was also demonstrated. As a final test, fluorescent lipid metabolites formed within cells loaded with BFLPIP2 were separated from a cell lysate as well as a single cell. This simple and

  4. Simulating the Osceola Mudflow Lahar Event in the Pacific Northwest using a GPU Based 2-Dimensional Hydraulic Model

    NASA Astrophysics Data System (ADS)

    Katz, B. G.; Eppert, S.; Lohmann, D.; Li, S.; Goteti, G.; Kaheil, Y. H.

    2011-12-01

    At 4,400 meters, Mount Rainer has been the point of origin for several major lahar events. The largest event, termed the "Osceola Mudflow," occurred 5,500 years ago and covered an area of approximately 550km2 with a total volume of deposited material from 2 to 4km3. Particularly deadly, large lahars are estimated to have maximum flow velocities in of 100km/h with a density often described as "Flowing Concrete." While rare, these events typically cause total destruction within a lahar inundation zone. It is estimated that approximately 150,000 people live on top of previous deposits left by lahars which can be triggered by anything from earthquakes to glacial and chemical erosion of volcanic bedrock over time to liquefaction caused by extreme rainfall events. A novel methodology utilizing a 2 dimensional hydraulic model has been implemented allowing for high resolution (30m) lahar inundation maps to be generated. The utility of this model above or in addition to other methodologies such as that of Iverson (1998), lies in its portability to other lahar zones as well as its ability to model any total volume specified by the user. The process for generating lahar flood plains requires few inputs including: a Digital Terrain Map of any resolution (DTM), a mask defining the locations for lahar genesis, a raster of friction coefficients, and a time series depicting uniform material accumulation over the genesis mask which is allowed to flow down-slope. Finally, a significant improvement in speed has been made for solving the two dimensional model by utilizing the latest in graphics processing unit (GPU) technology which has resulted in a greater than 200 times speed up in model run time over previous CPU-based methods. The model runs for the Osceola Mudflow compare favorably with USGS derived inundation regions as derived using field measurements and GIS based approaches such as the LAHARZ program suit. Overall gradation of low to high risk match well, however the new

  5. Hydraulic Modeling of Alluvial Fans along the Truckee Canal using the 2-Dimensional Model SRH2D

    NASA Astrophysics Data System (ADS)

    Wright, J.; Kallio, R.; Sankovich, V.

    2013-12-01

    Alluvial fans are gently sloping, fan-shaped landforms created by sediment deposition at the ends of mountain valleys. Their gentle slopes and scenic vistas are attractive to developers. Unfortunately, alluvial fans are highly flood-prone, and the flow paths of flood events are highly variable, thereby placing human developments at risk. Many studies have been performed on alluvial fans in the arid west because of the uncertainty of their flow paths and flood extents. Most of these studies have been focused on flood elevations and mitigation. This study is not focused on the flood elevations. Rather, it is focused on the attenuation effects of alluvial fans on floods entering and potentially failing a Reclamation canal. The Truckee Canal diverts water from the Truckee River to Lahontan Reservoir. The drainage areas along the canal are alluvial fans with complex distributary channel networks . Ideally, in nature, the sediment grain-size distribution along the alluvial fan flow paths would provide enough infiltration and subsurface storage to attenuate floods entering the canal and reduce risk to low levels. Human development, however, can prevent the natural losses from occurring due to concentrated flows within the alluvial fan. While the concentrated flows might mitigate flood risk inside the fan, they do not lower the flood risk of the canal. A 2-dimensional hydraulic model, SRH-2D, was coupled to a 1-dimensional rainfall-runoff model to estimate the flood attenuation effects of the alluvial fan network surrounding an 11 mile stretch of the Truckee Canal near Fernley, Nevada. Floods having annual exceedance probabilities ranging from 1/10 to 1/100 were computed and analyzed. SRH-2D uses a zonal approach for modeling river systems, allowing areas to be divided into separate zones based on physical parameters such as surface roughness and infiltration. One of the major features of SRH-2D is the adoption of an unstructured hybrid mixed element mesh, which is based

  6. Design and operation of a portable scanner for high performance microchip capillary array electrophoresis.

    PubMed

    Scherer, James R; Liu, Peng; Mathies, Richard A

    2010-11-01

    We have developed a compact, laser-induced fluorescence detection scanner, the multichannel capillary array electrophoresis portable scanner (McCAEPs) as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses. The instrument contains a confocal optical system with a rotary objective for detecting four different fluorescence signals, a pneumatic system consisting of two pressure/vacuum pumps and 28 individual addressable solenoid valves for control of on-chip microvalves and micropumps, four Polymerase Chain Reaction (PCR) temperature control systems, and four high voltage power supplies for electrophoresis. The detection limit of the instrument is ~20 pM for on-chip capillary electrophoresis of fluorescein dyes. To demonstrate the system performance for forensic short tandem repeat (STR) analysis, two experiments were conducted: (i) electrophoretic separation and detection of STR samples on a 96-lane microfabricated capillary array electrophoresis microchip. Fully resolved PowerPlex(®) 16 STR profiles amplified from 1 ng of 9947A female standard DNA were successfully obtained; (ii) nine-plex STR amplification, sample injection, separation, and fluorescence detection of 100-copy 9948 male standard DNA in a single integrated PCR- capillary electrophoresis microchip. These results demonstrate that the McCAEPs can be used as a versatile control and detection instrument that operates integrated microfluidic devices for high-performance forensic human identification. PMID:21133459

  7. Design and operation of a portable scanner for high performance microchip capillary array electrophoresis

    NASA Astrophysics Data System (ADS)

    Scherer, James R.; Liu, Peng; Mathies, Richard A.

    2010-11-01

    We have developed a compact, laser-induced fluorescence detection scanner, the multichannel capillary array electrophoresis portable scanner (McCAEPs) as a platform for electrophoretic detection and control of high-throughput, integrated microfluidic devices for genetic and other analyses. The instrument contains a confocal optical system with a rotary objective for detecting four different fluorescence signals, a pneumatic system consisting of two pressure/vacuum pumps and 28 individual addressable solenoid valves for control of on-chip microvalves and micropumps, four Polymerase Chain Reaction (PCR) temperature control systems, and four high voltage power supplies for electrophoresis. The detection limit of the instrument is ˜20 pM for on-chip capillary electrophoresis of fluorescein dyes. To demonstrate the system performance for forensic short tandem repeat (STR) analysis, two experiments were conducted: (i) electrophoretic separation and detection of STR samples on a 96-lane microfabricated capillary array electrophoresis microchip. Fully resolved PowerPlex® 16 STR profiles amplified from 1 ng of 9947A female standard DNA were successfully obtained; (ii) nine-plex STR amplification, sample injection, separation, and fluorescence detection of 100-copy 9948 male standard DNA in a single integrated PCR- capillary electrophoresis microchip. These results demonstrate that the McCAEPs can be used as a versatile control and detection instrument that operates integrated microfluidic devices for high-performance forensic human identification.

  8. [In situ photopolymerization of polyacrylamide-based preconcentrator on a microfluidic chip for capillary electrophoresis].

    PubMed

    Yamamoto, Sachio

    2012-01-01

    Microchip electrophoresis is widely used for microfluidics and has been studied extensively over the past decade. Translation of capillary electrophoresis methods from traditional capillary systems to a microchip platform provides rapid separation and easy quantitation of sample components. However, most microfluidic systems suffer from critical scaling problems. One promising solution to this problem is online sample preconcentration of all analytes in a sample reservoir before the separation channel. Herein, the following three techniques for online preconcentration during microchip electrophoresis are proposed: (1) in situ fabrication of an ionic polyacrylamide-based preconcentrator on a simple poly(methyl methacrylate) microfluidic chip for perm-selective preconcentration and capillary electrophoretic separation of anionic compounds, (2) simultaneous concentration enrichment and electrophoretic separation of weak acids on a microchip using an in situ photopolymerized carboxylate-type polyacrylamide gels as the perm-selective preconcentrator, and (3) microchip electrophoresis of oligosaccharides using lectin-immobilized preconcentrator gels fabricated by in situ photopolymerization. These techniques are expected to be powerful tools for clinical and pharmaceutical studies with on-line preconcentration during microchip electrophoresis. PMID:23023420

  9. Comparative analysis of excretory-secretory antigens of Trichinella spiralis and Trichinella britovi muscle larvae by two-dimensional difference gel electrophoresis and immunoblotting

    PubMed Central

    2012-01-01

    Background Trichinellosis is a zoonotic disease in humans caused by Trichinella spp. The present study was undertaken to discover excretory-secretory (E-S) proteins from T. spiralis and T. britovi muscle larvae (ML) that hold promise for species-specific diagnostics. To that end, the purified E-S proteins were analyzed by fluorescent two-dimensional difference gel electrophoresis (2-D DIGE) coupled with protein identification by liquid chromatography-tandem mass spectrometry (LC-MS/MS). To search for immunoreactive proteins that are specifically recognized by host antibodies the E-S proteins were subjected to two-dimensional (2-DE) immunoblotting with antisera derived from pigs experimentally infected with T. spiralis or T. britovi. Results According to 2-D DIGE analysis, a total of twenty-two proteins including potentially immunogenic proteins and proteins produced only by one of the two Trichinella species were subjected to LC-MS/MS for protein identification. From these proteins seventeen could be identified, of which many were identified in multiple spots, suggesting that they have undergone post-translational modification, possibly involving glycosylation and/or proteolysis. These proteins included 5'-nucleotidase, serine-type protease/proteinase, and p43 glycoprotein (gp43) as well as 49 kDa E-S protein (p49). Our findings also suggest that some of the commonly identified proteins were post-translationally modified to different extents, which in certain cases seemed to result in species-specific modification. Both commonly and specifically recognized immunoreactive proteins were identified by 2-DE immunoblotting; shared antigens were identified as gp43 and different protease variants, whereas those specific to T. britovi included multiple isoforms of the 5'-nucleotidase. Conclusions Both 2-D DIGE and 2-DE immunoblotting approaches indicate that T. spiralis and T. britovi produce somewhat distinctive antigen profiles, which contain E-S antigens with potential

  10. Improved solubilization of surface proteins from Listeria monocytogenes for 2-DE.

    PubMed

    Mujahid, Sana; Pechan, Tibor; Wang, Chinling

    2007-11-01

    Solubilization of bacterial surface (cell wall and membrane-associated) proteins for 2-DE is challenging, particularly in the case of Gram-positive bacteria. This is primarily due to strong protein association with the cell wall peptidoglycan and protein hydrophobicity. We solubilized surface proteins for 2-DE from the Gram-positive pathogen Listeria monocytogenes using mutanolysin, which digests cell wall peptidoglycan, and one of three different mixtures of zwitterionic detergent and chaotropes: (i) CHAPS/urea, (ii) amidosulfobetaine-14 (ASB-14)/urea/thiourea (iii) N-decyl-N,N'-dimethyl-3-ammonio-1-propanesulfonate/urea/thiourea. Cell lysis with mutanolysin followed by solubilization with ASB-14/urea/thiourea gave the highest overall protein yield with the best 2-DE resolution. Protein spot identification by MALDI-TOF/TOF-MS analysis revealed 29 characterized surface proteins of L. monocytogenes, 17 of which have not previously been reported on the surface proteome map. This is the first report describing the successful solubilization and 2-DE of L. monocytogenes proteins bound to the cell surface via an LPXTG motif or by a hydrophobic tail. The increase in surface proteome coverage obtained by mutanolysin and ASB-14/urea/thiourea solubilization suggests the utility of this method for future analytical and comparative studies of surface proteins from Listeria, and possibly other Gram-positive bacteria, using 2-DE proteomic analysis. An updated 2-DE reference map of L. monocytogenes surface proteins is presented. PMID:17922522

  11. Determination of preservatives in soft drinks by capillary electrophoresis with ionic liquids as the electrolyte additives.

    PubMed

    Sun, Bingbing; Qi, Li; Wang, Minglin

    2014-08-01

    A capillary electrophoresis method for separating preservatives with various ionic liquids as the electrolyte additives has been developed. The performances for separation of the preservatives using five ionic liquids with different anions and different substituted group numbers on imidazole ring were studied. After investigating the influence of the key parameters on the separation (the concentration of ionic liquids, pH, and the concentration of borax), it has been found that the separation efficiency could be improved obviously using the ionic liquids as the electrolyte additives and tested preservatives were baseline separated. The proposed capillary electrophoresis method exhibited favorable quantitative analysis property of the preservatives with good linearity (r(2) = 0.998), repeatability (relative standard deviations ≤ 3.3%) and high recovery (79.4-117.5%). Furthermore, this feasible and efficient capillary electrophoresis method was applied in detecting the preservatives in soft drinks, introducing a new way for assaying the preservatives in food products. PMID:24910409

  12. Application of capillary electrophoresis-electrospray ionization mass spectometry in the determination of molecular diversity.

    PubMed Central

    Dunayevskiy, Y M; Vouros, P; Wintner, E A; Shipps, G W; Carell, T; Rebek, J

    1996-01-01

    By means of capillary electrophoresis coupled online to electrospray ionization MS, a library of theoretically 171 disubstituted xanthene derivatives was analyzed. The method allowed the purity and makeup of the library to be determined: 160 of the expected compounds were found to be present, and 12 side-products were also detected in the mixture. Due to the ability of capillary electrophoresis to separate analytes on the basis of charge, most of the xanthene derivatives could be resolved by simple capillary electrophoresis-MS procedures even though 124 of the 171 theoretical compounds were isobaric with at least one other molecule in the mixture. Any remaining unresolved peaks were resolved by MS/MS experiments. The method shows promise for the analysis of small combinatorial libraries with fewer than 1000 components. PMID:8650235

  13. Protein electrophoresis as a diagnostic and prognostic tool in raptor medicine.

    PubMed

    Tatum, L M; Zaias, J; Mealey, B K; Cray, C; Bossart, G D

    2000-12-01

    Plasma proteins of 139 healthy adult birds of prey from 10 species were separated by electrophoresis to characterize and document normal reference ranges and species-specific electrophoretic patternsand to evaluate the value of this technique for health screening, disease diagnosis, and prognostic indication. Species studied included bald eagle (Haliaeetus leucocephalus), red-tailed hawk (Buteo jamaicensis), barn owl (Tyto alba), great horned owl (Bubo virginianus), turkey vulture (Cathartes aura), Harris' hawk (Parabuteo unicinctus), Stellar's sea eagle (Haliaeetus pelagicus), barred owl (Strix varia), screech owl (Otus asio), and black vulture (Coragyps atratus). Several clinical cases show the diagnostic/therapeutic value of protein electrophoresis in raptors. This study establishes species-specific reference ranges for several birds of prey and discusses the benefit of electrophoresis as a diagnostic technique in health screens, as a diagnostic aid in conjunction with other tests, and as a prognostic indicator in clinical evaluation of raptors. PMID:11428396

  14. DNA sequencing with capillary electrophoresis and single cell analysis with mass spectrometry

    SciTech Connect

    Fung, N.

    1998-03-27

    Since the first demonstration of the laser in the 1960`s, lasers have found numerous applications in analytical chemistry. In this work, two different applications are described, namely, DNA sequencing with capillary gel electrophoresis and single cell analysis with mass spectrometry. Two projects are described in which high-speed DNA separations with capillary gel electrophoresis were demonstrated. In the third project, flow cytometry and mass spectrometry were coupled via a laser vaporization/ionization interface and individual mammalian cells were analyzed. First, DNA Sanger fragments were separated by capillary gel electrophoresis. A separation speed of 20 basepairs per minute was demonstrated with a mixed poly(ethylene oxide) (PEO) sieving solution. In addition, a new capillary wall treatment protocol was developed in which bare (or uncoated) capillaries can be used in DNA sequencing. Second, a temperature programming scheme was used to separate DNA Sanger fragments. Third, flow cytometry and mass spectrometry were coupled with a laser vaporization/ionization interface.

  15. Continuous flow electrophoresis system experiments on shuttle flights STS-6 and STS-7

    NASA Technical Reports Server (NTRS)

    Snyder, Robert S.; Rhodes, Percy H.; Miller, Teresa Y.

    1988-01-01

    The development of a space continuous flow electrophoresis system (CFES) is discussed. The objectives of the experiment were: (1) to use a model sample material at a high concentration to evaluate the continuous flow electrophoresis process in the McDonnell Douglass CFES instrument and compare its separation resolution and sample throughput with related devices on Earth, and (2) to expand the basic knowledge of the limitations imposed by fluid flows and particle concentration effects on the electrophoresis process by careful design and evaluation of the space experiment. Hemoglobin and polysaccharide were selected as samples of concentration effects. The results from space show a large band spread of the high concentration of the single species of hemoglobin that was principally due to the mismatch of electrical conductivity between the sample and buffer.

  16. Use of capillary electrophoresis and indirect detection to quantitate in-capillary enzyme-catalyzed microreactions.

    PubMed

    Zhang, Y; el-Maghrabi, M R; Gomez, F A

    2000-04-01

    The use of capillary electrophoresis and indirect detection to quantify reaction products of in-capillary enzyme-catalyzed microreactions is described. Migrating in a capillary under conditions of electrophoresis, plugs of enzyme and substrate are injected and allowed to react. Capillary electrophoresis is subsequently used to measure the extent of reaction. This technique is demonstrated using two model systems: the conversion of fructose-1,6-bisphosphate to dihydroxyacetone phosphate and glyceraldehyde-3-phosphate by fructose-biphosphate aldolase (ALD, EC 4.1.2.13), and the conversion of fructose-1,6-bisphosphate to fructose-6-phosphate by fructose-1,6-bisphospatase (FBPase, EC 3.1.3.11). These procedures expand the use of the capillary as a microreactor and offer a new approach to analyzing enzyme-mediated reactions. PMID:10892022

  17. Two-dimensional fluorescence difference gel electrophoresis for comparative proteomics profiling

    PubMed Central

    Tannu, Nilesh S; Hemby, Scott E

    2007-01-01

    Quantitative proteomics is the workhorse of the modern proteomics initiative. The gel-based and MuDPIT approaches have facilitated vital advances in the measurement of protein expression alterations in normal and disease phenotypic states. The methodological advance in two-dimensional gel electrophoresis (2DGE) has been the multiplexing fluorescent two-dimensional fluorescence difference gel electrophoresis (2D-DIGE). 2D-DIGE is based on direct labeling of lysine groups on proteins with cyanine CyDye DIGE Fluor minimal dyes before isoelectric focusing, enabling the labeling of 2–3 samples with different dyes and electrophoresis of all the samples on the same 2D gel. This capability minimizes spot pattern variability and the number of gels in an experiment while providing simple, accurate and reproducible spot matching. This protocol can be completed in 3–5 weeks depending on the sample size of the experiment and the level of expertise of the investigator. PMID:17487156

  18. Preparative liquid column electrophoresis of T and B lymphocytes at gravity = 1

    NASA Technical Reports Server (NTRS)

    Van Oss, C. J.; Bigazzi, P. E.; Gillman, C. F.; Allen, R. E.

    1974-01-01

    Vertical liquid columns containing low-molecular-weight dextran density gradients can be used for preparative lymphocyte electrophoresis on earth, in simulation of zero gravity conditions. Another method that has been tested at 1 g, is the electrophoresis of lymphocytes in an upward direction in vertical columns. By both methods up to 100 million lymphocytes can be separated at one time in a 30-cm glass column of 8-mm inside diameter, at 12 V/cm, in two hours. Due to convection and sedimentation problems, the separation at 1 g is less than ideal, but it is expected that at zero gravity electrophoresis will probe to be a uniquely powerful cell separation tool.

  19. Studies with sample conductivity, insertion rates, and particle deflection in a continuous flow electrophoresis system

    NASA Technical Reports Server (NTRS)

    Williams, G., Jr.

    1982-01-01

    The continuous flow electrophoresis system makes electrophoresis possible in a free-flowing film of aqueous electrolyte medium. The sample continuously enters the electrolyte at the top of the chamber and is subjected to the action of a lateral dc field. This divides the sample into fractions since each component has a distinctive electrophoretic mobility. Tests were made using monodisperse polystyrene latex microspheres to determine optimum sample conductivity, insertion rates and optimum electric field applications as baseline data for future STS flight experiments. Optimum sample flow rates for the selected samples were determined to be approximately 26 micro-liters/min. Experiments with samples in deionized water yielded best results and voltages in the 20 V/cm to 30 V/cm range were optimum. Deflections of formaldehyde fixed turkey and bovine erythrocytes were determined using the continuous flow electrophoresis system. The effects of particle interactions on sample resolution and migration in the chamber was also evaluated.

  20. Evaluation of the Separability of Monodisperse Polystyrene Latex Microspheres in a Continuous Flow Electrophoresis System

    NASA Technical Reports Server (NTRS)

    Williams, G., Jr.

    1983-01-01

    The continuous flow electrophoresis system makes electrophoresis possible in a free flowing film of aqueous electrolyte medium. The sample is introduced at one end of the chamber and is subjected to a lateral dc field. This process separates the sample into fractions since each component has a distinctive electrophoric mobility. Evaluations were made of sample conductivity and buffer conductivity as they affect sample band spread and separation using the Continuous Particle Electrophoresis (CPE) system. Samples were prepared from mixtures of 5 percent and 1 percent polystyrene latex (PSL) microspheres which were .4, .56 and .7 microns in diameter. These were prepared in electrolyte media 1x and 3x the conductivity of the curtain buffer, approximately 150 and 450 micro mhos/cm. Samples with matched conductivities produced greater resolution and less band spread than those with 3x the conductivity of the curtain buffer.

  1. Enhancing resolution of free-flow zone electrophoresis via a simple sheath-flow sample injection.

    PubMed

    Yang, Ying; Kong, Fan-Zhi; Liu, Ji; Li, Jun-Min; Liu, Xiao-Ping; Li, Guo-Qing; Wang, Ju-Fang; Xiao, Hua; Fan, Liu-Yin; Cao, Cheng-Xi; Li, Shan

    2016-07-01

    In this work, a simple and novel sheath-flow sample injection method (SFSIM) is introduced to reduce the band broadening of free-flow zone electrophoresis separation in newly developed self-balance free-flow electrophoresis instrument. A needle injector was placed in the center of the separation inlet, into which the BGE and sample solution were pumped simultaneously. BGE formed sheath flow outside the sample stream, resulting in less band broadening related to hydrodynamics and electrodynamics. Hemoglobin and C-phycocyanin were successfully separated by the proposed method in contrast to the poor separation of free-flow electrophoresis with the traditional injection method without sheath flow. About 3.75 times resolution enhancement could be achieved by sheath-flow sample injection method. PMID:27121853

  2. A review of microdialysis coupled to microchip electrophoresis for monitoring biological events

    PubMed Central

    Saylor, Rachel A.; Lunte, Susan M.

    2015-01-01

    Microdialysis is a powerful sampling technique that enables monitoring of dynamic processes in vitro and in vivo. The combination of microdialysis with chromatographic or electrophoretic methods yields along with selective detection methods yields a “separation-based sensor” capable of monitoring multiple analytes in near real time. Analysis of microdialysis samples requires techniques that are fast (<1 min), have low volume requirements (nL–pL), and, ideally, can be employed on-line. Microchip electrophoresis fulfills these requirements and also permits the possibility of integrating sample preparation and manipulation with detection strategies directly on-chip. Microdialysis coupled to microchip electrophoresis has been employed for monitoring biological events in vivo and in vitro. This review discusses technical considerations for coupling microdialysis sampling and microchip electrophoresis, including various interface designs, and current applications in the field. PMID:25637011

  3. Single DNA electrophoresis in Pluronic F127 in a real-time fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    You, Seungyong; van Winkle, David

    2007-03-01

    Electrophoresis is the separation of bio-molecules in a sieving medium by applying an electric field. The Pluronic F127 gel was introduced as a new sieving medium for electrophoresis. The mobility of DNA in this gel is not fully explained by conventional reptation theories. Here, in our work, the migration of single DNA molecule pre-stained was studied on the gel electrophoresis by real-time fluorescence microscopy. Separations were performed on dsDNA fragments ranging in length from 200 base pairs (bp) to 2500 bp in pluronic gel in various concentrations. Evidence is presented that in some cases DNA fragments electrophorese along gel crystallite grain boundaries and in other cases directly through gel crystallites. This is direct observation of DNA migration through the pluronic gel on a microscopic scale.

  4. Diffusion of DNA during gel electrophoresis; a predictive function spanning the relevant regimes

    NASA Astrophysics Data System (ADS)

    McCormick, Laurette; Slater, Gary

    2004-03-01

    Gel electrophoresis is used extensively to separate DNA. Diffusion of the DNA bands during electrophoresis is an important phenomenon which reduces the resolution obtained. As with DNA mobility, the diffusion of DNA can be split into several different regimes, each described by relevant theory. Unfortunately, until recently there was no single formula for DNA mobility or diffusion that could be used in more than one regime. However, Van Winkle and co workers [Van Winkle DH, Beheshti A, Rill RL, ELECTROPHORESIS 23 (1): 15-19 JAN 2002] have successfully developed an analytical function to analyze DNA mobility data, throughout the relevant regimes. We present the development of a complementary function for the analysis of DNA diffusion. This function should be very useful both in analyzing DNA electrophoretic data, and as a predictive tool.

  5. Separating DNA with different topologies by atomic force microscopy in comparison with gel electrophoresis.

    PubMed

    Jiang, Yong; Rabbi, Mahir; Mieczkowski, Piotr A; Marszalek, Piotr E

    2010-09-23

    Atomic force microscopy, which is normally used for DNA imaging to gain qualitative results, can also be used for quantitative DNA research, at a single-molecular level. Here, we evaluate the performance of AFM imaging specifically for quantifying supercoiled and relaxed plasmid DNA fractions within a mixture, and compare the results with the bulk material analysis method, gel electrophoresis. The advantages and shortcomings of both methods are discussed in detail. Gel electrophoresis is a quick and well-established quantification method. However, it requires a large amount of DNA, and needs to be carefully calibrated for even slightly different experimental conditions for accurate quantification. AFM imaging is accurate, in that single DNA molecules in different conformations can be seen and counted. When used carefully with necessary correction, both methods provide consistent results. Thus, AFM imaging can be used for DNA quantification, as an alternative to gel electrophoresis. PMID:20799746

  6. Two-dimensional electrophoresis of plant proteins with phastsystem using nonequilibrium pH gradient separation.

    PubMed

    Ferullo, J M; Nespoulous, L

    1991-10-01

    We have adapted a two-dimensional electrophoretic technique described by P. Z. O'Farrell et al. (Cell 12, 1133-1142, 1977) to Phastsystem, resolving both acidic and basic proteins by using nonequilibrium pH gradient electrophoresis in the first dimension and sodium dodecyl sulfate polyacrylamide gel electrophoresis in the second dimension. Protein separation was optimized for the analysis of plant proteins. The use of the Phastsystem apparatus reduced times of preparation and separation, allowing the rapid screening of plant proteins on a large scale of isoelectric points. This technique was used for the immunodetection and characterization of two stress-induced proteins in irradiated tomato leaves. PMID:1789413

  7. Modified SSCP method using sequential electrophoresis of multiple nucleic acid segments

    DOEpatents

    Gatti, Richard A.

    2002-10-01

    The present invention is directed to a method of screening large, complex, polyexonic eukaryotic genes such as the ATM gene for mutations and polymorphisms by an improved version of single strand conformation polymorphism (SSCP) electrophoresis that allows electrophoresis of two or three amplified segments in a single lane. The present invention also is directed to new mutations and polymorphisms in the ATM gene that are useful in performing more accurate screening of human DNA samples for mutations and in distinguishing mutations from polymorphisms, thereby improving the efficiency of automated screening methods.

  8. Pulsed-field gel electrophoresis for epidemiologic studies of Campylobacter hyointestinalis isolates.

    PubMed Central

    Salama, S M; Tabor, H; Richter, M; Taylor, D E

    1992-01-01

    Campylobacter hyointestinalis was isolated from five members of the same family who had previously consumed raw milk. Pulsed-field gel electrophoresis of genomic DNAs from the five strains, after digestion with restriction endonuclease SalI, revealed that three strains had identical genome patterns and therefore appeared to be related, whereas the other two had completely different genome patterns and appeared to be unrelated. We report here for the first time the isolation of C. hyointestinalis from family members who had consumed raw milk. Our study also demonstrates the usefulness of pulsed-field gel electrophoresis for epidemiologic studies of this unusual campylobacter. Images PMID:1500503

  9. Proposal of New Rewritable Printing Media Using Electrophoresis and Confirmation of Its Mechanism

    NASA Astrophysics Data System (ADS)

    Hoshino, Yasushi; Ogura, Masahiro; Sano, Takayuki

    2004-10-01

    A new rewritable printing media using electrophoresis and selective heating is proposed to contribute to the reduction in paper consumption by printers. The mechanism is that when a heated part of the rewritable media is melted, white particles in that part of the media are able to move by electrophoresis. The media is initialized by heating its entire surface under the condition of voltage application and imaging is carried out by selective heating under the condition of an applied reversed-polarity voltage. Using a mixture system of carnauba wax and particles coated with titanium oxide (TiO2), the feasibility of the mechanism is confirmed.

  10. Physics and gel electrophoresis: using terminal velocity to characterize molecular weight

    NASA Astrophysics Data System (ADS)

    Viney, Christopher; Fenton, Richard A.

    1998-11-01

    Protein molecular weights are commonly characterized by gel electrophoresis. Biology textbooks typically quote an empirical, approximate relationship between migration rate and molecular weight, relying on an inappropriately simplistic model of spherical particles travelling at their terminal velocity through a viscous medium. We show how the model can be modified to derive a physically realistic equation that relates migration rate and molecular weight, and that mirrors experimentally observed behaviour. We suggest that gel electrophoresis provides an interesting interdisciplinary context in which to exercise several basic principles that are encountered through introductory physics courses. Finally, we provide additional examples of practical situations where the concept of terminal velocity can be elaborated and applied.

  11. The determination of molecular weights of biologically active proteins by cetyltrimethylammonium bromide-polyacrylamide gel electrophoresis.

    PubMed

    Akin, D T; Shapira, R; Kinkade, J M

    1985-02-15

    A novel cetyltrimethylammonium bromide-polyacrylamide gel electrophoresis system which is useful for the separation of native forms of proteins consistent with their molecular weights is reported here. Many proteins examined in this system demonstrated the same association patterns which have been shown by other techniques to exist under nondenaturing conditions. In addition, biological activity could be assayed directly in the gel after electrophoresis. Based on the peculiar characteristics of cetyltrimethylammonium bromide, a possible explanation which may account for the behavior of proteins in this system is presented. PMID:4003759

  12. Differentiation of opium and poppy straw using capillary electrophoresis and pattern recognition techniques.

    PubMed

    Reid, Raymond G; Durham, David G; Boyle, Susanne P; Low, Ann S; Wangboonskul, Jinda

    2007-12-12

    Opium samples from four different locations and poppy straw from different plant varieties have been assayed using micellar capillary electrophoresis incorporating a sweeping technique. Individual alkaloids (morphine, codeine, papaverine, noscapine, thebaine, oripavine, reticuline and narceine) were quantitatively determined in the different samples by a validated capillary electrophoresis method. Unsupervised pattern recognition of the opium samples and the poppy straw samples using hierarchical cluster analysis (HCA) and principal component analysis (PCA), showed distinct clusters. Supervised pattern recognition using soft independent modelling of class analogy (SIMCA) was performed to show individual groupings and allow unknown samples to be classified according to the models built using the CZE assay results. PMID:18022406

  13. Optimization of pulsed-field gel electrophoresis protocols for Salmonella Paratyphi A subtyping.

    PubMed

    Chen, Chunxia; Zhao, Yingwei; Han, Hui; Pang, Bo; Zhang, Jingyun; Yan, Meiying; Diao, Baowei; Cui, Zhigang; Zhou, Haijian; Liang, Weili; Feng, Yanfang; Kan, Biao

    2012-04-01

    Salmonella enterica serovar Paratyphi A infection has caused public health problems in some countries in recent years. Pulsed-field gel electrophoresis (PFGE) has been used for the subtyping and epidemiological investigations of some serotypes of Salmonella, mainly in outbreaks caused by non-typhoidal Salmonella. In this study, different restriction endonucleases and electrophoresis parameters were compared for the PFGE subtyping by using Salmonella Paratyphi A strain panels. Two protocols for the enzymes SpeI and XbaI showed higher discriminatory power, which may facilitate epidemiological analysis for more accurate case definition, and clonality study of Salmonella Paratyphi A. PMID:22443482

  14. pI-Control in Comparative Fluorescence Gel Electrophoresis (CoFGE) using amphoteric azo dyes.

    PubMed

    Hanneken, Marina; Šlais, Karel; König, Simone

    2015-06-01

    Amphoteric azo dyes were used for internal control of pI values in Comparative two-dimensional Fluorescence Gel Electrophoresis (CoFGE) [1]. The 2D-gel images of separated Escherichia coli proteins as well as those of colored amphoteric dyes separated by isoelectric focussing are presented. The latter were used to correct for variation in the first electrophoretic dimension and further improve protein coordinate assignment in 2D-gel electrophoresis. Data tables are supplied to demonstrate pI-value calibration and the effect on the assignment of protein spot coordinates. PMID:26217748

  15. pI-Control in Comparative Fluorescence Gel Electrophoresis (CoFGE) using amphoteric azo dyes

    PubMed Central

    Hanneken, Marina; Šlais, Karel; König, Simone

    2015-01-01

    Amphoteric azo dyes were used for internal control of pI values in Comparative two-dimensional Fluorescence Gel Electrophoresis (CoFGE) [1]. The 2D-gel images of separated Escherichia coli proteins as well as those of colored amphoteric dyes separated by isoelectric focussing are presented. The latter were used to correct for variation in the first electrophoretic dimension and further improve protein coordinate assignment in 2D-gel electrophoresis. Data tables are supplied to demonstrate pI-value calibration and the effect on the assignment of protein spot coordinates. PMID:26217748

  16. Should routine laboratories stop doing screening serum protein electrophoresis and replace it with screening immune-fixation electrophoresis? No quick fixes: Counterpoint.

    PubMed

    Smith, Joel D; Raines, Geoffrey; Schneider, Hans G

    2016-06-01

    Monoclonal gammopathies are characterised by the production of a monoclonal immunoglobulin or free light chains by an abnormal plasma cell or B-cell clone and may indicate malignancy or a precursor (MGUS). There is currently no consensus on the initial test or combination of tests to be performed in suspected monoclonal gammopathies but serum protein electrophoresis and urine protein electrophoresis are commonly requested as initial investigations. If abnormal, immunofixation electrophoresis is then performed to confirm the presence of paraprotein and to determine its heavy and light chain type. Recently, some groups have developed simplified "screening" IFE methods for use in parallel to SPEP for the detection monoclonal gammopathies. We argue here that screening IFE may be of benefit in clinical laboratories using SPEP with poor resolution in the β-region, assisting in the detection of mainly IgA paraprotein, but may be of less benefit in laboratories utilising higher resolution gels. Further it may increase the detection of trace bands of questionable clinical significance, representing transient phenomena in infectious and auto-immune conditions or very low risk MGUS. The increased detection of these bands using screening IFE would require further patient follow up, possibly causing unnecessary patient anxiety and additional follow up healthcare costs. PMID:26677889

  17. Development of polymeric coatings for control of electro-osmotic flow in ASTP MA-011 electrophoresis technology experiment

    NASA Technical Reports Server (NTRS)

    Patterson, W. J.

    1976-01-01

    The development of a methyl cellulose based coating system for control of electro-osmotic flow at the walls of electrophoresis cells is described. Flight electrophoresis columns were coated with this system, resulting in a flight set of six columns. In flight photography of MA-011 electrophoretic separations verified control of electro-osmotic flow.

  18. Investigation of capillary free-flow electrophoresis for separation of Co, Cr, and As species in aqueous solution

    SciTech Connect

    Ketterer, M.E.; Kozerski, G.E.; Ritacco, R.; Painuly, P.

    1997-01-01

    Application of a prototype capillary free-flow electrophoresis (CFFE) device for separation of different solution species of Co, Cr, and As is explored. A unique free-flow electrophoresis (FFE) design is employed, which makes use of internal capillary cooling tubes to minimize thermal convection due to Joule heating.

  19. Phenotyping breast cancer cell lines EM-G3, HCC1937, MCF7 and MDA-MB-231 using 2-D electrophoresis and affinity chromatography for glutathione-binding proteins

    PubMed Central

    2010-01-01

    Background Transformed phenotypes are common to cell lines derived from various cancers. Proteome profiling is a valuable tool that may reveal uncharacteristic cell phenotypes in transformed cells. Changes in expression of glutathione S-transferases (GSTs) and other proteins interacting with glutathione (GSH) in model cell lines could be of particular interest. Methods We compared the phenotypes of breast cell lines EM-G3, HCC1937, MCF7 and MDA-MB-231 using 2-D electrophoresis (2-DE). We further separated GSH-binding proteins from the cell lines using affinity chromatography with GSH-Sepharose 4B, performed 2-DE analysis and identified the main protein spots. Results Correlation coefficients among 2-DE gels from the cell lines were lower than 0.65, pointing to dissimilarity among the cell lines. Differences in primary constituents of the cytoskeleton were shown by the 2-D protein maps and western blots. The spot patterns in gels of GSH-binding fractions from primary carcinoma-derived cell lines HCC1937 and EM-G3 were similar to each other, and they differed from the spot patterns of cell lines MCF7 and MDA-MB-231 that were derived from pleural effusions of metastatic mammary carcinoma patients. Major differences in the expression of GST P1-1 and carbonyl reductase [NADPH] 1 were observed among the cell lines, indicating differential abilities of the cell lines to metabolize xenobiotics. Conclusions Our results confirmed the applicability of targeted affinity chromatography to proteome profiling and allowed us to characterize the phenotypes of four breast cancer cell lines. PMID:20731849

  20. Analysis of rRNA Gene Methylation in Arabidopsis thaliana by CHEF-Conventional 2D Gel Electrophoresis.

    PubMed

    Mohannath, Gireesha; Pikaard, Craig S

    2016-01-01

    Contour-clamped homogenous electric field (CHEF) gel electrophoresis, a variant of Pulsed-field gel electrophoresis (PFGE), is a powerful technique for resolving large fragments of DNA (10 kb-9 Mb). CHEF has many applications including the physical mapping of chromosomes, artificial chromosomes, and sub-chromosomal DNA fragments, etc. Here, we describe the use of CHEF and two-dimensional gel electrophoresis to analyze rRNA gene methylation patterns within the two ~4 million base pair nucleolus organizer regions (NORs) of Arabidopsis thaliana. The method involves CHEF gel electrophoresis of agarose-embedded DNA following restriction endonuclease digestion to cut the NORs into large but resolvable segments, followed by digestion with methylation-sensitive restriction endonucleases and conventional (or CHEF) gel electrophoresis, in a second dimension. Resulting products are then detected by Southern blotting or PCR analyses capable of discriminating rRNA gene subtypes. PMID:27576719

  1. Is capillary electrophoresis on microchip devices able to genotype uridine diphosphate glucuronosyltransferase 1A1 TATA-box polymorphisms?

    PubMed

    Minucci, Angelo; Canu, Giulia; De Bonis, Maria; Delibato, Elisabetta; Capoluongo, Ettore

    2014-06-01

    In this commentary, we focused our attention on capillary electrophoresis. It achieves the efficient separation of molecular species by the application of high voltages to samples in solution. Actually, capillary electrophoresis can be performed on microchip devices, based on an automated and miniaturized electrophoresis system, based on lab-on-a-chip technology. By this technology it is possible to separate nucleic acid fragments (DNA or RNA) with respect to sizing accuracy and sizing resolution. Currently, two automated capillary electrophoresis on microchips devices are available: the Agilent 2100 Bioanalyzer and the Experion™ Automated Electrophoresis System. In this study, we evaluated if the CE is able to distinguish the three uridine diphosphate glucuronosyltransferase 1A1 TATA-box genotypes. PMID:24687976

  2. ANALYSIS OF ANIONIC METALLIZED AZO AND FORMAZAN DYES BY CAPILLARY ELECTROPHORESIS/MASS SPECTROMETRY

    EPA Science Inventory

    Capillary electrophoresis-mass spectrometry was applied to the separation of several anionic dyes containing copper(II), chromium(III), or cobalt(III) as part of the dye molecule. The dyes were separated using a 110 cmX50 mu m uncoated fused-silica capillary and a 5 mM ammonium a...

  3. [Characterization of Acholeplasma Strains by Horizontal Polyacrylamide Flat Gel Electrophoresis (author's transl)].

    PubMed

    Boden, K; Kirchhoff, H

    1977-01-01

    Proteins extracted with phenol-acetic acid-water (2:1:0.5, w/v/v) from Acholeplasma laidlawii (PG 8), A. granularum (BTS-39), A. oculi (19L), A. modicum (PG 49), A. axanthum (S743) and the Acholeplasma strains C1 and C112 (which were isolated from aborted horse foetuses) were compared by electrophoresis in horizontal acidic polyacylamide flat gel using the electrophoresis equipment LKB Multiphor 2117. In this system the gels are not prepared in the electrophoresis chamber but between glas plates. For electrophoresis they are applied onto a special cooling plate. This makes it possible to produce a number of identical gels (from the same gel mixture and polymerized under the same conditions) what can be important for comparing investigations. The gels can be stored for more than 4 weeks in the refrigerator at +4 degrees C. Marked differences were observed between the electrophoretic patterns of each of the established species and the horse strains C1 and C112. The results are in agreement with those obtained in serological investigations in which the strains C1 and C112 were different from the established Acholeplasma species. PMID:848216

  4. Understanding Electrophoresis through the Investigation of Size, Shape, and Charge of pH Indicators

    ERIC Educational Resources Information Center

    Brenner, Ryan K.; Hess, Kenneth R.; Morford, Jennifer L.

    2015-01-01

    A laboratory experiment was designed for upper-level students in a Chemical Analysis course to illustrate the theoretical and practical applications of 0.8% agarose gel electrophoresis and to reinforce an understanding of weak acids/bases using easy-to-visualize pH indicators. The careful choice of indicators included acid and base types with…

  5. The Gel Electrophoresis Markup Language (GelML) from the Proteomics Standards Initiative

    PubMed Central

    Gibson, Frank; Hoogland, Christine; Martinez-Bartolomé, Salvador; Medina-Aunon, J. Alberto; Albar, Juan Pablo; Babnigg, Gyorgy; Wipat, Anil; Hermjakob, Henning; Almeida, Jonas S; Stanislaus, Romesh; Paton, Norman W; Jones, Andrew R

    2011-01-01

    The Human Proteome Organisation’s Proteomics Standards Initiative (HUPO-PSI) has developed the GelML data exchange format for representing gel electrophoresis experiments performed in proteomics investigations. The format closely follows the reporting guidelines for gel electrophoresis, which are part of the Minimum Information About a Proteomics Experiment (MIAPE) set of modules. GelML supports the capture of metadata (such as experimental protocols) and data (such as gel images) resulting from gel electrophoresis so that laboratories can be compliant with the MIAPE Gel Electrophoresis guidelines, while allowing such data sets to be exchanged or downloaded from public repositories. The format is sufficiently flexible to capture data from a broad range of experimental processes, and complements other PSI formats for mass spectrometry data and the results of protein and peptide identifications to capture entire gel-based proteome workflows. GelML has resulted from the open standardisation process of PSI consisting of both public consultation and anonymous review of the specifications. PMID:20677327

  6. Capillary electrophoresis of the mycotoxin zearalenone using cyclodextrin-enhanced fluorescence

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Certain of the cyclodextrins are capable of significantly enhancing the native fluorescence of the estrogenic mycotoxin zearalenone (ZEN). Twenty-two cyclodextrins (CDs) were screened for their ability to enhance the fluorescence of ZEN in a capillary electrophoresis-laser induced fluorescence (CE-...

  7. Microfabricated capillary electrophoresis chip and method for simultaneously detecting multiple redox labels

    DOEpatents

    Mathies, Richard A.; Singhal, Pankaj; Xie, Jin; Glazer, Alexander N.

    2002-01-01

    This invention relates to a microfabricated capillary electrophoresis chip for detecting multiple redox-active labels simultaneously using a matrix coding scheme and to a method of selectively labeling analytes for simultaneous electrochemical detection of multiple label-analyte conjugates after electrophoretic or chromatographic separation.

  8. PRECONCENTRATION OF ALIPHATIC AMINES FROM WATER DETERMINED BY CAPILLARY ELECTROPHORESIS WITH INDIRECT UV DETECTION

    EPA Science Inventory

    Preconcentration methodology based on adsorption chromatographies for enriching aliphatic amines (c1 to C4 substituted primary, secondary, and tertiary) and alkanolamines in water was studied by free zone capillary electrophoresis (CZE)with indirect UV detection. The solid-phase ...

  9. Aligning Goals, Assessments, and Activities: An Approach to Teaching PCR and Gel Electrophoresis

    ERIC Educational Resources Information Center

    Phillips, Allison R.; Robertson, Amber L.; Batzli, Janet; Harris, Michelle; Miller, Sarah

    2008-01-01

    Polymerase chain reaction (PCR) and gel electrophoresis have become common techniques used in undergraduate molecular and cell biology labs. Although students enjoy learning these techniques, they often cannot fully comprehend and analyze the outcomes of their experiments because of a disconnect between concepts taught in lecture and experiments…

  10. Capillary Electrophoresis Analysis of Cations in Water Samples: An Experiment for the Introductory Laboratory

    ERIC Educational Resources Information Center

    Pursell, Christopher J.; Chandler, Bert; Bushey, Michelle M.

    2004-01-01

    Capillary electrophoresis is gradually working its way into the undergraduate laboratory curriculum. Typically, experiments utilizing this newer technology have been introduced into analytical or instrumental courses. The authors of this article have introduced an experiment into the introductory laboratory that utilizes capillary electrophoresis…

  11. Capillary electrophoresis of alkali and alkaline-earth cations with imidazole or benzylamine buffers

    SciTech Connect

    Morin, P.; Francois, C.; Dreux, M. . Lab. de Chimie Bioorganique et Analytique)

    1994-01-01

    The separation of alkali, alkaline earth, and ammonium cations in several samples of water was achieved by capillary electrophoresis with indirect UV detection. A solution of imidazole (10[sup [minus]2] M, pH 4.5) was used as a buffer to resolve a mixture of six cations (K[sup +], Na[sup +], Ca[sup 2+], Ba[sup 2+], Li[sup +] and Mg[sup 2+]) by capillary electrophoresis at 214 nm in less than 10 min. The addition of potassium cation to the running buffer has an influence on the resolution of Ca[sup 2+]/Na[sup +] and Na[sup +]/Mg[sup 2+] peaks. A linear relationship between the corrected peak area and concentration was obtained in the 1--10 ppm range for these cations using a hydrodynamic injector. This electrophoretic system permitted the separation of these inorganic cations at a 50 ppb-level concentration with a hydrodynamic injection, thus making it possible to quantitatively determine their presence in mineral waters by capillary electrophoresis. At pH 4.5, potassium and ammonium unfortunately have identical ionic mobilities causing them to comigrate in an imidazole buffer. Using an alkaline solution of benzylamine as carrier electrolyte, their separation can be successfully achieved with excellent resolution at 204 nm. The analyses of tap water and several mineral waters have been achieved by capillary electrophoresis.

  12. Determination of the specific activities of methionine sulfoxide reductase A and B by capillary electrophoresis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A capillary electrophoresis (CE) method for the determination of methionine sulfoxide reductase A and methionine sulfoxide reductase B activities in mouse liver is described. The method is based on detection of the 4-(dimethylamino)azobenzene-4’-sulfonyl derivative of L-methionine (dabsyl Met), the ...

  13. ESTIMATION OF HRW WHEAT HEAT DAMAGE BY DSC, CAPILLARY ZONE ELECTROPHORESIS, PHOTOACOUSTIC SPECTROSCOPY AND RHEOMETRY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of heat damage was estimated using Hard Red Winter (HRW) wheat varieties grown in Oklahoma. The testing was done on wheat kernels, flour, and isolated starch. Whole-wheat kernels were analyzed by Photoacoustic Spectroscopy (PAS). Flour was analyzed by DSC, Capillary Electrophoresis (CE...

  14. Means and method for capillary zone electrophoresis with laser-induced indirect fluorescence detection

    DOEpatents

    Yeung, Edwards; Kuhr, Werner G.

    1991-04-09

    A means and method for capillary zone electrphoresis with laser-induced indirect fluorescence detection. A detector is positioned on the capillary tube of a capillary zone electrophoresis system. The detector includes a laser which generates a laser beam which is imposed upon a small portion of the capillary tube. Fluorescence of the elutant electromigrating through the capillary tube is indirectly detected and recorded.

  15. Disposable pen-shaped capillary gel electrophoresis cartridge for fluorescence detection of bio-molecules

    NASA Astrophysics Data System (ADS)

    Amirkhanian, Varoujan; Tsai, Shou-Kuan

    2014-03-01

    We introduce a novel and cost-effective capillary gel electrophoresis (CGE) system utilizing disposable pen-shaped gelcartridges for highly efficient, high speed, high throughput fluorescence detection of bio-molecules. The CGE system has been integrated with dual excitation and emission optical-fibers with micro-ball end design for fluorescence detection of bio-molecules separated and detected in a disposable pen-shaped capillary gel electrophoresis cartridge. The high-performance capillary gel electrophoresis (CGE) analyzer has been optimized for glycoprotein analysis type applications. Using commercially available labeling agent such as ANTS (8-aminonapthalene-1,3,6- trisulfonate) as an indicator, the capillary gel electrophoresis-based glycan analyzer provides high detection sensitivity and high resolving power in 2-5 minutes of separations. The system can hold total of 96 samples, which can be automatically analyzed within 4-5 hours. This affordable fiber optic based fluorescence detection system provides fast run times (4 minutes vs. 20 minutes with other CE systems), provides improved peak resolution, good linear dynamic range and reproducible migration times, that can be used in laboratories for high speed glycan (N-glycan) profiling applications. The CGE-based glycan analyzer will significantly increase the pace at which glycoprotein research is performed in the labs, saving hours of preparation time and assuring accurate, consistent and economical results.

  16. Nanometal-Oxide Sunscreen Agents by Capillary Electrophoresis-Inductively Coupled Plasma Mass Spectrometry

    EPA Science Inventory

    Capillary electrophoresis with detection by ICPMS is being explored to characterize nanomaterials in waste water treatment plant effluents. TiO2 and ZnO, being widely used as UV filters in personal care products, plastics, and paints, are of concern as they enter the environment...

  17. Characterization of IHSS Natural Organic Matter by Capillary Electrophoresis and Flourescence Spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Capillary electrophoresis (CE) and fluorescence spectroscopy have recently been used in natural organic matter (NOM) studies. In this study, we characterized fulvic acids (5), humic acids (6) and unprocessed NOM (2) samples obtained from the International Humic Substances Society (IHSS) using these ...

  18. Capillary Electrophoresis and Fluorescence Excitation-Emission Matrix Spectroscopy for Characterization of Humic Substances

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Capillary electrophoresis (CE) and fluorescence spectroscopy have been used in natural organic matter (NOM) studies. In this study, we characterized five fulvic acids, six humic acids and two unprocessed NOM samples obtained from the International Humic Substances Society (IHSS) using these two ana...

  19. CAPILLARY ELECTROPHORESIS-ELECTROSPRAY MASS SPECTRA OF THE HERBICIDES PARAQUAT AND DIQUAT

    EPA Science Inventory

    The positive ion electrospray mass spectra of the quaternary ammonium salt herbicides paraquat and diquat are examined by on-line separation with capillary electrophoresis (CE) and by direct infusion of the analytes. The analytes are separated by CE in 7-10 min at pH 3.9 in 50% m...

  20. Means and method for capillary zone electrophoresis with laser-induced indirect fluorescence detection

    DOEpatents

    Yeung, Edward S.; Kuhr, Werner G.

    1996-02-20

    A means and method for capillary zone electrphoresis with laser-induced indirect fluorescence detection. A detector is positioned on the capillary tube of a capillary zone electrophoresis system. The detector includes a laser which generates a laser beam which is imposed upon a small portion of the capillary tube. Fluorescence of the elutant electromigrating through the capillary tube is indirectly detected and recorded.