bluuebunny/crossref_metadata_2025_split huggingface dataset:
Capillary electrophoresis of rat liver microsomes in polymer solutions
Authors: Radko Sergey P. , Sokoloff Alexander V. , Garner Mark M. , Chrambach Andreas 
Published: May 2005

AbstractRat liver microsome components were separated by capillary zone eletrophoresis in buffers containing substituted agarose, agarose crosslinked polyacrylamide, polyacrylamide, polyethylene glycol and dextran of various molecular weights. The best resolution of the components was obtained with polymers of 10–21 nm geometric mean radius. Both the crude and the purified preparations of microsomes exhibit a single major peak. In a Tris‐borate‐EDTA (TBE) buffer, containing polyacrylamide of 5 × 106 molecular weight, it has a retardation coefficient, KR, of 0.77 × 0.02. Translation of the KR value to geometric mean radii, R, on the basis of the standard curve applicable to polymer solutions, KR vs. R, with polystyrene carboxylate size standards in both dextran and polyacrylamide solution allows one to estimate a value of R as 13–16 nm for the major microsome component. The value is smaller than expected from electron microscopic measurements (100–250 nm), possibly due to the chemical and geometric differences between microsomes and the polystyrene particles used as size standards. The crude preparation also contains a minor component which is smaller and less charged than the major component. Another component, apparently very much larger than the major one, is seen in TBE buffer but not in a potassium‐N‐(2‐hydroxyethyl)piperazine‐2′‐(2‐ethanesulfonic acid) buffer and is therefore thought to be an artifact of interaction with borate. After a short incubation under conditions promoting delayed microsome fusion, i.e. in presence of GTP and Mg++ and in the absense of polyethylene glycol, the electrophoretic pattern changes dramatically: it now exhibits five unretarded highly mobile and, therefore, presumbly large components in addition to the two original retarded components of the microsome and a less highly charged species similar in KR to the smaller of the original two components.
Capillary electrophoresis of human serum proteins and apolipoproteins
Authors: Lehmann Rainer , Liebich Hartmut , Grübler Gerald , Voelter Wolfgang 
Published: May 2005

AbstractCapillary electrophoresis (CE) analysis of human serum proteins and apolipoproteins is described and compared with sodium dodecyl sulfate‐polyacryl‐amide gel electrophoresis and cellulose acetate membrane electrophoresis. Under optimized CE conditions apolipoproteins I could be determined directly in the serum and quantitated with a linear correlation between peak area and concentration up to a concentration of 100 mg/dL.
Visualization of barley β‐glucan degrading isozymes after gel isoelectric focusing
Authors: Menteur Sophie , Jestin Louis , Risacher Thierry , Branlard Gérard 
Published: May 2005

AbstractA simple method to study the polymorphism of barley (Hordeum vulgare L.) β‐(1–3, 1–4)‐glucanases (specific enzymes of barley β‐glucan hydrolysis) is described. Proteins of a crude extract of germinated barley kernels were separated in an immobilized pH gradient in two pH ranges (pH 3–10.5 and pH 4–7). β‐glucanases were visualized by contact printing with a polyacrylamide gel containing β‐glucan or lichenan. Patterns of β‐glucanases were revealed by staining with Congo Red with resultant clear zones on a stained background. Various conditions of germination, extraction and visualization were investigated. New isozyme bands could be detected and their nature and origin discussed.
Transient bisalbuminemia: Separation by isoelectric focusing of human albumin fractions linked to different numbers of benzylpenicilloyl groups
Authors: Rocha Jorge , Bohner Jürgen , Kömpf Jost 
Published: May 2005

AbstractThe behavior of modified human serum albumin (HSA) from penicillin‐treated patients and in vitro conjugated samples was studied by isoelectric focusing in the absence and presence of 8 M urea. It is shown that samples classified as bisalbuminemic after separation under native conditions display a spectrum of several major bands with decreasing pIs when focusing is done in the presence of 8 M urea. In this spectrum, the distances between two consecutive major bands are as expected from a 2‐unit charge difference. Since this result is compatible with the blockage of a lysine residue with a benzylpenicilloyl (BPO) group, each major band can be interpreted as representing an HSA fraction covalently linked to different numbers of benzylpenicilloyl groups. The separation methods presented therefore seem to constitute simple and reliable tools for a detailed assessment of the extent of BPO covalent binding to HSA both in vivo and in vitro.
Two‐dimensional electrophoresis of proteins: An updated protocol and implications for a functional analysis of the genome
Authors: Klose Joachim , Kobalz Ursula 
Published: May 2005

AbstractThe two‐dimensional electrophoresis (2‐DE) technique developed by Klose in 1975 (Humangenetik 1975, 26, 211–234), independently of the technique developed by O'Farrell (J. Biol. Chem. 1975, 250, 4007–4021), has been revised in our laboratory and an updated protocol is presented. This protocol is the result of our experience in using this method since its introduction. Many modifications and suggestions found in the literature were also tested and then integrated into our original method if advantageous. Gel and buffer composition, size of gels, use of stacking gels or not, necessity of isoelectric focusing (IEF) gel incubation, freezing of IEF gels or immediate use, carrier ampholytes versus Immobilines, regulation of electric current, conditions for staining and drying the gels – these and other problems were the subject of our concern. Among the technical details and special equipment which constitute our 2‐DE method presented here, a few features are of particular significance: (i) sample loading onto the acid side of the IEF gel with the result that both acidic and basic proteins are well resolved in the same gel; (ii) use of large (46 × 30 cm) gels to achieve high resolution, but without the need of unusually large, flat gel equipment; (iii) preparation of ready‐made gel solutions which can be stored frozen, a prerequisite, among others, for high reproducibility. Using the 2‐DE method described we demonstrate that protein patterns revealing more than 10 000 polypeptide spots can be obtained from mouse tissues. This is by far the highest resolution so far reported in the literature for 2‐DE of complex protein mixtures. The 2‐DE patterns were of high quality with regard to spot shape and background. The reproducibility of the protein patterns is demonstrated and shown to be thoroughly satisfactory. An example is given to show how effectively 2‐DE of high resolution and reproducibility can be used to study the genetic variability of proteins in an interspecific mouse backcross (Mus musculus × Mus spretus) established by the European Backcross Collaborative Group for mapping the mouse genome. We outline our opinion that the structural analysis of the human genome, currently pursued most intensively on a worldwide scale, should be accompanied by a functional analysis of the genome that starts from the proteins of the organism.
Two‐dimensional polyacrylamide gel electrophoresis with immobilized pH gradients in the first dimensin (IPG‐Dalt): The state of the art and the controversy of vertical versus horizontal systems
Authors: Görg Angelika , Boguth GÜNther , Obermaier Christian , Posch Anton , Weiss Walter 
Published: May 2005

AbstractAfter having established the basic protocol of two‐dimensional electrophoresis with immobilized pH gradients in the first dimension (IPG‐Dalt) in 1988 (A. Görg et al., Electrophoresis 1988, 9, 531–546), some critical parameters of the actual IPG‐Dalt protocols as well as the results obtained with horizontal and vertical second‐dimensional sodium dodecyl sulfate‐electrophoresis are demonstrated and discussed.
Sequence analysis of wheat grain allergens separated by two‐dimensional electrophoresis with immobilized pH gradients
Authors: Posch Anton , Weiss Walter , Wheeler Colin , Dunn Michael J. , Görg Angelika 
Published: May 2005

AbstractMicropreparative two‐dimensional (2‐D) gel electrophoresis with immobilized pH gradients (4–8) in the first dimension (IPG‐DALT) was optimized for the separation of salt‐soluble wheat grain proteins associated with bakers' asthma disease. The resolved polypeptides were electroblotted onto a polyvinylidene difluoride (PVDF) membrane and incubated with the pooled sera from four asthmatic bakers. Bound IgE was demonstrated by alkaline phosphatase conjugated anti‐human IgE. Major IgE binding was detected in the 27 kDa, 37 kDa and, to a lesser extent, in the 14–18 kDa area of the 2‐D immunoblots, respectively. Since the main purpose of our study was to determine the N‐terminal amino acid sequences of the major wheat grain allergens, N‐terminal sequencing was performed for six out of a total of eleven major allergens located in the 27 kDa area, for one out of two 37 kDa allergens, and for two out of four 14–18 kDa allergens. Our results revealed that two of the 27 kDa polypeptides are clearly related to several Acyl‐CoA oxidase variants of barley and rice, whereas no significant homologies were found for the remaining four 27 kDa allergens analyzed. The N‐terminus of the 37 kDa allergen appeared to be blocked so that no sequence information was obtained, while the two 14–18 kDa allergens analyzed were identified as members of the wheat α‐amylase‐inhibitor family.
Inside SWISS‐2DPAGE database
Authors: Sanchez Jean‐Charles , Appel Ron D. , Golaz Olivier , Pasquali Christian , Ravier Florence , Bairoch Amos , Hochstrasser Denis F. 
Published: May 2005

AbstractSeveral two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) databases have been established and updated for more than 15 years. Only recently have developments of computer networks and high‐speed transfer protocols provided the required tools for sharing comprehensive and hypermedia 2‐D PAGE databases. This publication describes the SWISS‐2DPAGE database structure. Proteins present in samples of human tissue, cells, cell lines and body fluids are assembled and described in an accessible uniform format. SWISS‐2DPAGE can be freely accessed through the World‐Wide Web (WWW) network on the ExPASy molecular biology server.
Rapid detection of the main human plasma glycoproteins by two‐dimensional polyacrylamide gel electrophoresis lectin affinoblotting
Authors: Golaz Olivier , Gravel Patricia , Walzer Claude , Turler Hans , Balant Luc , Hochstrasser Denis F. 
Published: May 2005

AbstractGlycoprotein modifications in the glycan moiety can occur in diseases such as cancers, inflammatory processes and alcoholism. We combined high‐resolution two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) with lectin affinoblotting in order to establish the normal human plasma glycoprotein map. Human plasma proteins were separated by mini 2‐D PAGE (7 × 9 cm), transferred onto polyvinylidene difluoride membranes and incubated with biotinylated lectins. We focused our study on lectins binding sialic acid and galactose residues. Known plasma glycoproteins such as αl‐antitrypsin, αl‐antichymotrypsin, α2‐HS glycoprotein, αl‐acid glycoprotein, haptoglobin β‐chain and transferrin were easily detected in ng amounts. This protocol was adequate to establish a normal plasma glycoprotein map and will allow the study of glycoproteins in diseases.
Effects of cyclosporine A on the rat liver and kidney protein pattern, and the influence of vitamin E and C coadministration
Authors: Benito Begoña , Wahl Daniel , Steudel Nicola , Cordier André , Steiner Sandra 
Published: May 2005

AbstractThe effects of cyclosporine A (CsA), a potent immunosuppressive drug, were examined in rat liver and kidney samples using two‐dimensinal electrophoretic protein analysis. Of a total of 370 liver and 336 kidney spots analyzed, 8% (29 spots) and 6% (19 spots), respectively, showed a significant drug‐induced change (p < 0.01), which was predominantly reflected in increased protein abundance (62% and 74% of the changes, respectively). Of the 48 proteins changed in either organ, 14 were most probably common to both tissues and one of these was significantly increased in both the liver and the kidney. Most of the other 13 showed similar trends (either increases or decreases) in both organs. However, the most striking drug effect seen in this study concerned an unindentified protein present only in the kidney, which completely disappeared upon CsA treatment. It was also investigated whether the drug‐induced changes could be prevented by the coadministration of the radical scavengers vitamin E and C with CsA. Spots changed by the administration of the drug were classified according to three different categories, based on their response profiles in rats treated with CsA in combination with the vitamins: (i) spots which were changed by CsA as well as by CsA in combination with the vitamins (12 liver and 4 kidney spots), (ii) spots which were changed by CsA and showed an additional increase of this change by CsA plus the vitamins (no liver and 4 kidney spots), and (iii) spots which were changed by CsA but not by CsA in combination with the vitamins (8 liver and 6 kidney spots). These results showed that in both organs the vitamins were able to prevent around 30% of the effects caused by CsA, and that two‐dimensional gel electrophoresis is an excellent tool to demonstrate such drug interactions at the molecular level.
Microheterogeneity of mouse antidextran monoclonal antibodies
Authors: Mimura Yusuke , Kabat Elvin A. , Tanaka Tatehiko , Fujimoto Masanori , Takeo Kazusuke , Nakamura Kazuyuki 
Published: May 2005

AbstractMouse antidextran monoclonal antibodies showed microheterogeneity which was analyzed by two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE). Not only the heavy (H) chains but also the light (L) chains were heter ogeneous in terms of isoelectric point (pI). The higher the pI, the more prominent the H chain spots. To demonstrate the cause of the microheterogeneity an IgG1 monoclonal antibody (mAb 35.8.2H) was examined especially for involvement of the sugar moiety in the microheterogeneity. The glycosylated region was determined in the Fc portion from serine 239 to methionine 309 by a glycan detection method using mild periodate oxidation, which confirms that the sugar chain is attached to the conserved glycosylation site of asparagine 297. However, charge heterogeneity of the H chain was not entirely attributed to the Fc because the papain digest of the antibody was separated into two Fc spots, a few Fd spots and two L chain spots by 2‐D PAGE. This indicates that factors other than the sugar moiety are responsible for charge heterogeneity of IgG monoclonal antibody. On the other hand, the H chain isoforms of lower pI were shown to be more susceptible to V8 protease by peptide mapping. This result strongly suggests the occurrence of deamidation at glutamine or asparagine residues.
Distance indices in a comparison between the A, D, I and R genomes of the Triticeae tribe
Authors: Zivy Michel , Madidi Saïd El , Thiellement Hervé 
Published: May 2005

AbstractTwo‐dimensional gel electrophoresis of shoot proteins was used to study the relationships between Triticum, Secal, and Hordeum. A high level of polymorphism was found among the 1275 spots scored: only 198 spots were found common to all. But, under the hypothesis that only allelic variations were observed, the mean number of alleles per locus was only two. Phenograms were built from different distance indices. All of them showed Triticum genomes A and D close to each other, Hordeum, far from the Triticum cluster, and Secale at an intermediate position. A discussion on the use of various distance indices is presented.
Electrophoretic, chromatographic and immunological studies of human urinary proteins
Authors: Büeler Martin R. , Wiederkehr Felix , Vonderschmitt Dieter J. 
Published: May 2005

AbstractUrinary proteins from both sexes were analyzed by high resolution two‐dimensional gel electrophoresis (2‐DE). For well reproducible 2‐DE patterns, the samples were concentrated and desalted in one step by vacuum dialysis. A reference map for urine proteins was established by the analysis of urine from 10 healthy persons. Proteins in urine that share immunogenicity with serum proteins were identified by use of antibody to whole human‐serum protein in an affinity‐column fractionation of urine and differential analysis of the adsorbed (serum component) and unadsorbed (non‐serum component) fractions. For identification of individual proteins, coelectrophoresis, immunoblotting and affinity chromatography with corresponding antibodies were used. Proteins identified in the map, besides known serum proteins, included: the subunit of Tamm‐Horsefall protein, the secretory component of IgA, constant breakdown products of αl‐antitrypsin and retinol‐binding protein, the five isoforms of the β chain of human chorionic gonadotropin and the subunit of prostatic acid phosphatase. In addition, we could demonstrate three proteins which are markedly pronounced in female urine, especially pregnant women. To get more information about the native properties of various urinary proteins, they were separated into four main peaks according to their sizes using fast protein liquid chromatography equipment. Possible interpolypeptide disulfide bonds were studied using a nonreducing 2‐DE system. 2‐DE in combination with other methods seems to be a valuable tool for the characterization of urinary proteins in defined renal or extra‐renal diseases. An example is given by analyzing the immune complexes from seven patients with a urinary tract infection.
Electrophopretic mobility of soft particles
Authors: Ohshima Hiroyuki 
Published: May 2005

AbstractA recent theory of electrophoresis of “soft” particles, i.e., polyelectrolytecoated colloidal particles (Ohshima, J. Colloid Interface Sci. 1994, 163, 474–483), is reviewed. This theory unites two different electrophoresis theories for rigid colloidal spherical particles and for spherical polyelectrolytes. An example is given of the analysis of electrophoretic mobility data on hydrogel‐layer‐coated latex particles.
A computer model for time‐based simulation of electrophoresis systems with freely defined initial and boundary conditions
Authors: Schafer‐Nielsen Claus 
Published: May 2005

AbstractA PC‐based program has been developed that allows the user to perform one‐dimensional computer simulations of electrophoresis systems with freely definable initial‐ and boundary conditions. The program can handle n constituents with n pK‐values and calculates constituent concentrations and derived parameters as a function of time. Results are displayed graphically on screen and are stored as data files for graphical hard‐copy processing.
Sensitive quantification of proteins by electrophoresis in gels by use of chemiluminescence
Authors: Vesterberg Olof , Acevedo Fernando , Bayard Cecilia 
Published: May 2005

AbstractFor protein quantitation in gels or blotting membranes, chemiluminescence (CL) offers the advantages of a substantial improvement of detection limits. Easy‐to‐use CL‐chemicals and specific probes such as antibodies conjugated to enzymes, e.g. alkaline phosphatase (AP) may be used in combination with a newly developed luminometer. CL was found to have low detection limits and a linear relation between relative light units (RLU) and the concentration of the antibody enzyme complex present over a wide concentration range. Measurements of the immunoglobulin IgE in dot blots and in blots after sodium dodecyl sulfate (SDS) electrophoresis under nonreducing conditions in agarose gels are described.
Electrophoretic size separation of proteins treated with sodium dodecyl sulfate in 1% agarose gels
Authors: Acevedo Fernando , Marín Viviana , Wasserman Moisés 
Published: May 2005

AbstractSeparation of proteins treated with sodium dodecyl sulfate (SDS) according to molecular size was achieved by discontinuous electrophoresis in vertical low‐concentration agarose gels. A linear relationship was found between the migration distance and the square root of the molecular weight. This holds for proteins in the range of 7–200 kDa separated in 1.25% w/v agarose gel slabs (7 × 7 × 0.15 cm) with 0.1% w/v SDS and sulfate as leading ion. The linear regression coefficient was 0.998. The molecular weight and charge of coions influenced the separation. Small ions with low pKa values were found suitable as coions. The migration distance of proteins treated with SDS varied linearly with the agarose concentration of the gel. The agarose type and quality affected the resolution of the SDS‐protein bands. We conclude that agarose gels can substitute polyacrylamide gels for the separation of proteins treated with SDS. A homogeneous agarose gel at a concentration of about 1% is a nonsieving support for electrophoresis. Therefore, the separation described here cannot be explained by the pore size of the gel. The results suggest that the separation is mainly due to the relative migration velocities of the coion and the proteins treated with SDS.
Rapid typing of variable number of tandem repeat locus in the human apolipoprotein B gene for DNA diagnosis of heart disease by polymerase chain reaction and capillary electrophoresis
Authors: Baba Yoshinobu , Tomisaki Riyo , Sumita Chinuyo , Morimoto Izumi , Sugita Sanae , Tsuhako Mitsutomo , Miki Tetsuro , Ogihara Toshio 
Published: May 2005

AbstractThe apolipoprotein B (apoB) variable number of tandem repeat (VNTR) alleles containing larger repeat units is a risk factor for coronary heart disease. Capillary electrophoresis (CE) in entangled polymer solution was applied to the analysis of polymerase chain reaction (PCR) amplified apoB VNTR locus for DNA diagnosis of heart disease. The CE separation gives an excellent resolution of two alleles differing by one or two 16 bp repeat units in the DNA size range up to 600 bp with high speed. The apoB alleles differing in length by 2 or 4 repeat units are readily distinguishable by CE in the DNA size range from 600 to 1000 bp. The plate number achieved was 1 million plates per meter. CE combining with PCR provides an excellent technique for accurate determination of the number of repeat units of apoB VNTR alleles and differentiation of heterozygous from homozygous individuals. Using the CE technique, the apoB VNTR loci from some individuals in genotyping were examined towards precise DNA diagnosis for coronary heart disease.
Influence of electrolyte composition on the effective electric field strength in capillary zone electrophoresis
Authors: Jumppanen Juho H. , Riekkola Marja‐Liisa 
Published: May 2005

AbstractIn capillary zone electrophoresis the electrophoretic velocity of an ion decreases as the concentration of the background electrolyte solution is increased. This is caused by changes in the electrophoretic mobility of the ion (μep) as well as by changes in the net force affecting it, namely the effective electric field strength (Eeff). The electrophoretic mobility of an ion is altered through changes in absolute viscosity of the electrolyte solution and changes in the solvated size of the ion. Eeff is altered mainly by changes in the magnitude of the charge asymmetry effect and the electrophoretic effect, both of which retard the motion of ions. In this study, the three‐marker technique was used to study the effect of background electrolyte concentration (0.02–0.08 M 3‐[cyclohexylamino]‐1‐propanesulfonic acid and counter ion (Li, Na, K, and Cs) on Eeff. It was found that the concentration of the background electrolyte markedly affects Eeff and that Eeff approaches E as the concentration of the background electrolyte approaches zero. The counter ion had a minor effect on Eeff: as the size of the hydrated radius of the counter ion increased, Eeff decreased. The three‐marker technique proved to be efficient for such determinations.