bluuebunny/crossref_metadata_2025_split huggingface dataset:
Electrophoretic characterization of esterase‐19 (ES‐19), a new arylesterase of the house mouse (Mus musculus)
Authors: Münz Michael , von Deimling Otto 
Published: May 2005

AbstractEsterase‐19 (ES‐19) of the house mouse, following specific visualization in electrophoretic and isoelectric focusing gels, is described. A list of diagnostic features was compiled including substrate specificity, inhibition and activation characteristics, temporal and spatial patterns of expression, distribution among laboratory and wild mouse strains. ES‐19 was classified as arylesterase (E.C. 3.1.1.2) and compared with ES‐16 and ES‐25, two other mouse arylesterases. Homology with brain esterases of rat, guinea pig, rabbit and man was found, and a uniform disegnation “arylesterase I” is proposed.
Reducing background interference in silver stained two‐dimensional polyacrylamide gels
Authors: Amess Robert , Fox John E. , Spragg Peter S. 
Published: May 2005

AbstractAmino acid analysis shows that commercial grades of agarose contain peptides. The quantity varies with the purity of the agarose. These findings agree with our observations that the extent of the background after silver‐staining for agarose gels or where agarose was used in the system (as in two‐dimensional polyacrylamide gel electrophoresis) was determined mainly by the quality of agarose used.
Induction of a mitochondrial matrix protein in human fibroblasts by interferon or poly I:C
Authors: Leigh Anderson N. 
Published: May 2005

AbstractThe abundance of protein Mitcon: 5, a major component of monocytes and granulocytes as observed by high‐resolution two‐dimensional electrophoresis, is increased in human fibrolasts treated with interferon or poly I:C. Production of this polypeptide is eliminated in cells treated with dinitrophenol, but not in cells treated with chloramphenicol, indicating that it is a cytoplasmically synthesized mitochondrial protein. Neither chloroquine nor tunicamycin, both of which interfere with proper targeting of lysozomal enzymes, affects Mitcon: 5 production. Actinomycin D prevents induction, indicating that new transcription is required. Cell fractionation studies show that Mitcon: 5 is found in the mitochondria/lysosome fraction, and that it is released from this fraction by treatments designed to liberate matrix enzymes. Mitcon: 5 thus appears to be the first example of a direct effect of interferon treatment on the composition of mitochondria. Furher characterization of this effect may help to explain side‐effects of interferon treatment, including fever.
A simplified procedure for evaluation and storage of isoelectric focusing gels prior to second‐dimension electrophoresis
Authors: Gower David J. , Tytell Michael 
Published: May 2005

AbstractA simple method of stain fixation of polyacrylamide isoelectric focusing tube gels is described. Gel handling and storage are simplified by fixation and second‐dimension quality is markedly improved over that from the original procedure involving quick freezing and frozen storage of the gel.
Relationship between electrophoretic mobility and activation of human monocytes
Authors: Bauer Johann , Hannig Kurt 
Published: May 2005

AbstractIsolated human monocytes showed an increased electrophoretic mobility (EM) during incubation in vitro. However, when the monocytes were isolated after activation in a mixed leukocyte culture, they tended to maintain their original EM during culturing. Activated and non‐activated monocytes were compared with respect to their activities during lymphocyte transformation. The lymphocytes which were used to test the activities of both kinds of monocytes had been depleted of all accessory cells by a two‐step physical isolation procedure including coutercurrent centrifugal elutriation and cell electrophoresis. Non‐activated monocytes supported concanavalin A (Con A) ‐ induced transformation of the purified lymphocytes optimally when the culture medium was supplemented by fetal calf serum (FCS). If, however, activated monocytes were used in lymphocyte transformation tests, Con A induced optimal [14C]thymine‐deoxyribose incorporation, even if FCS was replaced by autologous human plasma. The results suggest that there is a relationship between the stability of the EM of human monocytes and their ability to support lymphocyte proliferation during an in vivo immune response, when FCS is obviously not present.
Serum protein analysis on immobilized pH gradients with in situ adsorption of albumin on Dextran Blue
Authors: Gianazza Elisabetta , Giacon Patrizia , Astrua‐Testori Silvia , Righetti Pier Giorgio 
Published: May 2005

AbstractDextran Blue incorporated in concentrations up to 1% w/v into a gel layer buffered with Immobiline chemicals to pH 9.1 does not interfere with isoelectric focusing within an adjacent immobilized pH gradient (IPG) and retains its ability to strongly bind human serum albumin. With minor modifications of the procedure, it is possible to use Dextran Blue shelves in connection with either non‐denaturing or denaturing (i. e. containing 8 M urea) analysis media. Examples include easier typing of group‐specific component in an ultranarrow IPG against a completely clear background and improved resolution of transferrin and hemopexin in two‐dimensional separations.
Detection of non‐heme iron proteins following polyacrylamide gel electrophoresis
Authors: Ohmori Daijiro , Tanaka Yukiko , Yamakura Fumiyuki , Suzuki Koji 
Published: May 2005

AbstractNon‐heme iron proteins were stainned in the gel with bathophenanthroline disulfonate after polyacrylamide gel electrophoresis. This staining method proved more resistant to color fading than iron detection by α,α′‐dipyridyl, with >50% of the color being retained after 24 h by three ferredoxins of different origin. A linear relationship was found for the amount of iron in non‐heme iron proteins, applied to the gel, and the areas of densitometric tracings.
A combination of Coomassie Blue and concanavalin A ‐ based glycoprotein stains avoids ambiguities in the assignment of bands
Authors: Spatz Hanns‐Christof , Waehneldt Thomas V. , Neuhoff Volker 
Published: May 2005

AbstractA simple procedure is described to double‐stain sodium dodecyl sulfate‐polyacrylamide gels both with Coomassie Blue and by the concanavalin A ‐ horseradish peroxidase method to detect glycoproteins among the protein bands. Double staining proves useful in cases where many bands are present on the gels and where a comparison of banding patterns from separate gels or even separate lanes of the same gel cannot be carried out with sufficient accuracy.
Sensitive Nitro Blue Tetrazolium staining of proteins in high resolution two‐dimensional gel electrophoresis
Authors: Bahrman Nasser , Thiellement Hervé 
Published: May 2005

AbstractA new method is described for staining proteins after high resolution two‐dimensional gel electrophoresis. The polyacrylamide gel matrix is stained blue, by photoreduction of Nitro Blue Tetrazolium, while the spots of polypeptides remain unstained. The method is much more sensitive than Coomassie Blue staining and its sensitivity is close to that obtained by silver staining.
The effects of high salt concentrations in the samples on molecular weight determination in sodium dodecyl sulfate polyacrylamide gel electrophoresis
Authors: See Yew Phew , Olley Peter M. , Jackowski George 
Published: May 2005

AbstractFor proper molecular weight determination in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), the protein should bind the same amount of SDS. High ionic strength had previously been shown to reduce the amount of SDS bound to proteins, suggesting that high salt concentrations in the samples may interfere with molecular weight determination. We examined the effects of NaCl from 0 to 1 M in the samples on molecular weight determination of proteins in SDS‐PAGE. In our conditions of electrophoresis, the retardation coefficients (KR), free mobilities (Yo) and Rf values for proteins (Mr 20 000–66 000) are not affected by NaCl in the samples up to 0.8 M. Therefore molecular weight of these proteins can be determined by both the Ferguson plot and Rf vs. log Mr methods in the presence of NaCl in the samples up to 0.8 M and compared with standard protein samples in the absence of NaCl. The Rf, KR and Yo values for phosphorylase (Mr 97 400) are affected by NaCl from 0.4 M or higher in the samples. Preliminary results show that increasing sodium phosphate, pH 7.0 from 0.01 M to 0.1 M in the sample affect molecular weight determinations. The effects of phosphate are greater in the Ferguson plot than in the Rfvs. log Mr plot procedure. In the latter method, the effects are dependent on the acrylamide gel concentrations and on the size of the proteins. We conclude that the molecular weight of proteins (Mr 20 000 to 97 000) can be determined in the presence of high salt concentration in the sample if the unknown and standard protein samples contain the same amount of salt and buffer ions.
Improved classification of alpha‐1‐antitrypsin in immobilized pH gradients containing sucrose
Authors: Pascali Vincenzo L. , Conte Gabriella 
Published: May 2005

AbstractTwo narrow immobilized pH gradients (pH 4.35–4.85 and pH 4.40–4.80) were formulated by linear interpolation from standard 1 pH unit ranges. Both gradients proved excellent for alpha‐1‐antitrypsin phenotype separation. Gel casting was simplified and the properties of the gels during the run improved by replacing glycerol with sucrose in the polymerization solutions.
Electrophoretic transport equations: Electrophoretic models based on migration only and their interrelationships
Authors: Thormann Wolfgang , Mosher Richard A. 
Published: May 2005

AbstractThe general equations which describe the electrophoretic transport of components in solution are restated using Newman's general concept of mobilities. A concise derivation of the moving boundary equation and the regulating function ω from the continuity equation is presented. Various other regulating principles across moving and stationary boundaries are also discussed, which permits a review of the features and interrelationshpis of the electrophoretic models based on electromigration only. The effect of considering an interactive (dissociating) solvent on the mathematical treatment is discussed.
Two‐dimensional fractionation of some particular human DNA restriction fragments
Authors: Guérin Patrick , Lucotte Gérard 
Published: May 2005

AbstractA procedure which allows the resolution of a portion of the human genome into discrete bands corresponding to particular DNA fragments is described. Genomic DNA has been digested with a restriction endonuclease, and the resulting DNA fragments are fractionated according to size on a polyacrylamide gel. After elution of DNA each fraction is digested with a second restriction endonuclease and analysed after radioactive labeling on a second polyacrylamide gel. Autoradiograms of these gels reveal numerous bands with sizes below two kilobases. A reproducible pattern of bands between individuals tissues and species, is generated; most of them correspond to multicopy sequences, being cut simultaneously by these two particular restriction enzymes.
A discussion of the constraints which lead to steady state electrophoretic boundaries
Authors: Mosher Richard A. , Thormann Wolfgang 
Published: May 2005

AbstractThe electric field variation across moving boundaries in three component systems is analyzed with a model based on electromigration only (moving boundary equation and regulating function ω). A decrease in the electric field strength across a boundary in the direction of migration is the most general requirement for the establishment of steady state migrating boundaries. The formation of that electric field gradient is shown to be based on two fundamental constraints. These are (i) the higher net mobility of the leading component compared to that of the terminating constituent and (ii) the higher concentration ratio of the leading component to the terminating component ahead of the moving boundary compared to that in the terminating zone. The relations are discussed with respect to strong and weak electrolytes where all three components are present on either side of the boundary. The predictions are verified experimentally. Limitations of the model based on migration only are discussed.
Concentration of cerebrospinal fluid does not affect immunoglobulin G oligoclonal banding patterns
Authors: Shapshak Paul , Tourtellotte Wallace W. , Lee Mike M. , Staugaitis Susan M. , Cowan Tina , Ingram Tim 
Published: May 2005

AbstractThe effects of concentration on immunoglobulin G (IgG) oligoclonal bands after isoelectric focusing in cerebrospinal fluids (CSF) were examined using six concentration procedures, and the percent recoveries of IgG varied. The procedures, % IgG recoveries, and corresponding degrees of concentration are as follows: (1) Amicon CF25 membrane cone, 75 %, 45 ×; (2) Amicon CF50A membrane cone, 65 %, 3 ×; (3) Amicon CS15 Minicon, 47 %, 80 ×; (4) Microprodicon, 55 %, 35 ×; (5) collodion bag negative pressure dialysis, 50 %, 16 ×; and (6) microcollodion bag in Permasorb, 40–80 %, 10–60 ×. Ultracentrifugation of CSF at 100 000 g for 1 h prior to concentration using CF 25 membrane cones introduced not more than 5 % variation in the IgG concentrations. Isoelectric focusing was performed on 20 CSF samples concentrated by CF25 membrane cone, CS15 Minicon, and collodion bag, and no effects on the oligoclonal IgG banding patterns were found.
Background estimation in one‐dimensional electropherograms of whole‐cell protein extracts
Authors: Lasters Ignace , Leyns Frederik , Jackman Peter J. H. 
Published: May 2005

AbstractIn electropherograms of whole‐cell protein extracts a background absorbance due to many partially overlapping bands is usually observed. Since this background is in general not relevant for the identification of a given pattern, its removal will increase the discrimination between patterns, especially when the distance between patterns is measured on a correlation basis. Here we present an algorithm for the estimation of background envelope. The algorithm selects a set of concave kernels of the total profile and subsequently performs a Fourier smoothing of the trace linking these kernels. A fraction of this smoothed trace is then the background estimate. The method is illustrated for the analysis of total protein extracts of bacterial strains.
Characterisation of a glycoprotein in oviductal fluid by two‐dimensional electrophoresis and lectin binding to protein gel blots
Authors: Sutton Rosemary , Wallace Alan L. C. , Nancarrow Colin D. 
Published: May 2005

AbstractTwo‐dimensional electrophoresis and Western blotting with lectins were used to provide information about a glycoprotein without requiring its previous purification. The oestrus‐associated glycoprotein from ovine oviductal fluid has an isoelectric point of 4.7 and a subunit molecular weight (Mr) range of 70 000–90 000. Under non‐denaturing conditions, it is present as a high molecular weight aggregate or polymer although at low pH some monomer is also detected. The surface carbohydrate on this protein includes fucose, galactose and N‐acetyl galactosamine residues but not glucose, mannose or sialic acid.
A high resolution, rapid procedure for alpha 1‐antitrypsin phenotyping
Authors: Budowle Bruce , Murch Randall S. 
Published: May 2005

AbstractAn ultrathin‐layer polyacrylamide gel isoelectric focusing method is described for typing alpha 1‐antritrypsin. This method, which uses Pharmalyte pH 4.2–4.9 and a 9.5 cm or 7.0 cm inter‐electrode wick distance, clearly resolves the M1, M2, M3, S, Z and a number of rare variant allelic products. In addition, a rapid screening method for the detection of the Z, the most deficient variant, using a gel with an electrode wick distance of 5.0 cm, is also efficaceous in the detection of this and other rare allelic products.
Mechanical stabilization of large pore acrylamide gels by addition of linear polyacrylamide
Authors: Mills Nathaniel C. , Ilan Judith 
Published: May 2005

AbstractLinear polyacrylamide has been used to increase the tensile strength and stability of low percentage acrylamide gels in the range of 2.5 to 6.0 % w/v acrylamide. This added strength allows handling and staining of the gel without the excessive difficulty which is normally incurred when working in this range. Addition of up to 0.6 % w/v of linear polyacrylamide gives no pore size reduction. While addition of linear polyacrylamide to acrylamide gels using bisacrylamide as a crosslinker has a hazy appearance, the use of diallyltartardiamide as a crosslinker gives the desired clarity without loss of large pore size and improves adherence to glass walls. A further advantage of acrylamide gel matrices is that the traditional methods of silver staining can be used for detection of DNA. Thus, adding linear polyacrylamide to an acrylamide gel using diallytartardiamide as a crosslinker allows easy use of low percentage acrylamide gels for separation and detection of DNA fragments in the range of 70 to 2500 base pairs.
Improved esterase D typing on ultrathin isoelectric focusing gels using narrow interval Pharmalyte carrier ampholytes (pH 4.5–5.4) in conjunction with a separator and a thickness modified pH gradient
Authors: Gill Peter 
Published: May 2005

AbstractThree common alleles are found in esterase D: EsD 1, EsD 2 and EsD 5. In addition, a number of rare variants are found. The isoeiectric points of the EsD 1 and EsD 2 isoenzymes differ by only 0.02 pH units, which could lead to possible confusion between the two using conventional isoelectric focusing (IEF). Therefore, in order to ensure adequate separation of these two bands, it is necessary to use a narrow pH gradient. The separation of esterase D (EsD) has been improved on electrofocusing gels by incorporating a narrow range Pharmalyte (pH 4.5–5.4) in conjunction with the separator N‐(2‐hydroxyethyl)piperazine‐N′‐2‐ethanesulphonic acid (HEPES). This mixture resulted in the formation of a narrow pH range of pH 4.85–5.35 (the major typing bands of EsD fall within the range of pI 4.9–5.2). Resolution was further improved by the use of thickness modified pH gradients in ultrathin gels which flattened the pH gradient in the range of pH 5.0–5.1. The techniques used were shown to be effective for typing bloodstains up to 1 month old.
Sodium dodecyl sulfate agarose gel electrophoresis: Stacking of the samples in a short polyacrylamide layer
Authors: Rhalem Abdelkébir , Pery Pierre 
Published: May 2005

AbstractA metod is described for obtaining a sample‐stacking effect before sodium dodecyl sulfate‐agarose gel electrophoresis. The method requires no novel equipment and uses a discontinuous polyacrylamide‐agarose horizontal slab gel system. With the buffers selected, a complex mixture of proteins can be separated, according to the molecular weight of their subunits, in 15 % w/w agarose gels. The new gel system is versatile and may offer advantages over sodium dodecyl sulfate ‐ polyacrylamide gel electrophoresis in such applications as protein elution or enzyme renaturation.
An immunochemical method for detecting an inactive enzyme (bisphosphoglyceromutase) after Cellogel electrophoresis
Authors: Calvin Marie‐Claude , Préhu Marie‐Odette , Kechemir Dalila , Villeval Jean‐Luc , Rosa Raymonde 
Published: May 2005

AbstractA new method is described for detecting inactive enzymes or other proteins after Cellogel electrophoresis. The proteins were blotted from the Cellogel strip to a nitro‐cellulose membrane and then the blot was blocked before incubation in a specific antibody solution. The antigen‐antibody complex was located by a second antibody staining procedure employing peroxidase‐linked goat anti‐rabbit IgG. The method described is sensitive and could be of general use for the detection of proteins after Cellogel electrophoresis.
Identification of rat liver carboxylesterase isozymes (EC 3.1.1.1) using polyacrylamide gel electrophoresis and isoelectric focusing
Authors: Simon Babette , de Looze Stephen , Ronai Adam , von Deimling Otto 
Published: May 2005

AbstractThe banding pattern of rat liver carboxylesterases (EC 3.1.1.1) was demonstrated following polyacrylamide gel electrophoresis and isoelectric focusing using standardized conditions. Phenotypic variations, occurring in commonly used inbred rat strains, were compared. Separate isozymes were identified using genetic nomenclature. Individual bands were labelled; their electrophoretic parameters were estimated. Three hitherto genetically undefined zones were described and preliminarily classified as carboxylesterase isozymes. A scheme was provided to enable identification of liver esterases in rat strains not investigated in the present study.
Two‐dimensional gel analysis of urine proteins after acidified‐acetone extraction
Authors: Guevara Juan G. , Herbert Barbara H. , Martin Barbara A. 
Published: May 2005

AbstractAn effective, reproducible and simple method for separating proteins from other urine constituents is described. Approximately 95 % of detectable urine protein is collected in the void volume peak from a single molecular exclusion gel column following an acidified acetone extraction of lyophilized urine. Analysis of the final urine protein preparation, using two‐dimensional (2‐D) gel electrophoresis and staining with silver showed patterns comparable to those reported by others. An estimated 700 individual proteins were discernible in 2‐D electropherograms of 75 μg of urine proteins. This is a reliable procedure that makes possible the isolation of proteins excreted in urine, which can then be used in other types of analysis or in further purification of urinary antigens.
Suitability of sulfur labelling reagent (Amersham) as an in vitro protein radiolabelling reagent for two‐dimensional electrophoresis
Authors: Rabilloud Thierry , Therre HéLène 
Published: May 2005

AbstractThis report describes a method of in vitro labelling of proteins designed for subsequent analysis of the sample by two‐dimensional electrophoresis. As much as 106 cpm can be incorporated in 5 μg of proteins. As the reagent reacts with the amino groups, virtually all the proteins are labelled. Thus, this method is far more sensitive than silver staining and may prove useful when extremely low amounts of sample are to be analysed.
Computer simulation of immobilized pH gradients at acidic and alkaline extremes: A quest for extended pH intervals
Authors: Mosher Richard A. , Bier Milan , Righetti Pier Giorgio 
Published: May 2005

AbstractComputer simulations of the concentration profiles of simple biprotic ampholytes with Δ pKs 1, 2 and 3, on immobilized pH gradients (IPG) at extreme pH values (pH 3–4 and pH 10–11) show markedly skewed steady‐state profiles with increasing kurtosis at higher Δ pK values. Across neutrality all the peaks are symmetric irrespective of their Δ pK values, but they show very high contribution to the conductivity of the background gel and significant alteration of the local buffering capacity. The problems of skewness, due to the exponential conductivity profiles at low and high pHs, and of gel burning due to a strong electroosmotic flow generated by the net charges in the gel matrix, also at low and high pHs, are solved by incorporating in the IPG gel a strong viscosity gradient. This is generated by a gradient of linear polyacrylamide which is trapped in the gel by the polymerization process.