摘要:

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.

ISSN:0173-0835    

DOI:10.1002/elps.1150061202

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractThe electrophoretic properties of the four phosphoglucomutase (PGM1) allozymes were studied in the presence of different buffer constituents at pH values from 4.6 to 6.0. A separation of the allozymes allowing the identification of the 10 common phenotypes was obtained in the entire pH range. The separation of the “1+” and “1−” allozymes and of the “2+” and “2−” allozymes was poor near pH 6 but improved considerably at more acidic pH values. This indicated that the charge difference between the “1+” and the “1−” allozymes and between the “2+” and “2−” allozymes is due to substitution of a neutral amino acid in the “1+” and “2+” allozymes with histidine in the “1−” and “2−” allozymes. Above pH 5 the sequence from cathode to anode was “1−”,“1+”,“2−”,“2+” and below pH 5 it was “1−”,“2−”,“1+”,“2+”. At the lower pH values the separation between the “1−” and the “2−” allozymes and between the “1+” and the “2+” allozymes diminished. Hence only two PGM1species may exist at more acidic pH (pH« 4.6): a “+” enzyme and a “−” enzyme. The presence of di‐ and polyvalent carboxylic acids in the electrophoretic buffer reduced the mobility of the enzymes towards the cathode or changed it into a migration towards the anode. This “buffer effect” was greatest with the polyvalent carboxylic acids. It is suggested that it is caused by an adsorption of carboxylate ions onto the enzyme surface, which results in a transfer of negative charges to the enzyme. The “buffer effect” varied slightly from one enzyme species to another, and this may explain the general expe

ISSN:0173-0835    

DOI:10.1002/elps.1150061203

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractLow voltage isoelectric focusing (IEF) for the classification of esterase D (ESD) has been developed. This new phenotyping method is based on non‐equilibrium IEF. The obtained patterns might be isotachophoretic because the ESD bands were concentrated sharply and the values of their isoelectric point (pI) were somewhat higher than those found by equilibrium IEF. The method proved to be fast, easy and reliable for the differentiation of the six phenotypes determined by three codominant alleles, ESD*1, ESD*2 and ESD*7. The distribution of ESD phenotypes in Western Japan has been investigated and a new rare variant, ESD Yamaguchi (ESD Y), characterized by a slightly cathodal mobility to ESD 2, could be identified. The frequencies for the three alleles ESD*1, ESD*2 and ESD*7 were 0.600, 0.389 and 0.011, respectivel

ISSN:0173-0835    

DOI:10.1002/elps.1150061204

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractUsing the ISO‐DALT system for two‐dimensional (2‐D) electrophoresis and the TYCHO system for computer analysis of the resulting protein maps, we obtained high quality quantitative protein abundance data from Coomassie Brilliant Bluestained gels of mouse liver samples. High resolution gels allow more than 100 proteins to be measured with coefficients of variation less than 15 %. A comparison of results from two mouse strains (C57BL/6 and BALB/c) and the cross between them (BCF1) shows that a large number of qutive polymorphisms can be detected, and that, as expected, the amount of protein produced in the heterozygote is intermediate between the parental values. The system described is shown to be capable of reliably detecting decreases in protein abundance such as those expected to result from radiation‐induced deletion of one copy of a gene. The implications of these results for the study of gene regulation are discussed in relation to applications in genetics, toxicology, and differen

ISSN:0173-0835    

DOI:10.1002/elps.1150061205

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractTwo‐dimensional (2‐D) maps obtained with horizontal or vertical electrophoretic systems are compared. There are no differences in resolution, spot size and distribution, but significant differences in handling, Horizontal 2‐D electrophoresis, where the equilibrated isoelectric focusing (IEF) gel strip with plastic backing is applied to the surface of the horizontal sodium dodecyl sulfate (SDS)‐gel, is extremely simple and easy to perform. Due to the optimal flat‐to‐flat contact of the two gels, no size or edge trimming of the gels, no agarose overlays nor embedding gels are needed. The influence of IEF, performed with carrier ampholytes, Immobilines or immobilized pH gradients (IPG) with carrier ampholytes, on 2‐D maps is demonstrated. More protein has migrated into the IPG gel with carrier ampholytes, displaying an increased nu

ISSN:0173-0835    

DOI:10.1002/elps.1150061206

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

AbstractHuman peripheral blood lymphocytes were isolated from healthy donors and cultivated in the absence and presence of phytohemagglutinin (PHA) or pokeweed mitogen (PWM). Cell lysates of unlabelled lymphocytes or following stimulation of [H]leucine metabolically labelled lymphocytes were subjected to two‐dimensional electrophoresis. Differences in the protein patterns are described which are related to the method applied for visualization of the proteins (silver staining or fluorography), to the culture conditions (unstimulated, PHA or PWM stimulated lymphocytes) and to individual variations. Comparative analysis of the resulting patterns on silver stained gels and fluorograms revealed three qualitative and several quantitative differences. Comparing the two‐dimensional patterns of unstimulated and mitogenstimulated lymphocytes with silver staining, we recognized the quantitative increase of 6 protein spots after stimulation. Whereas protein patterns of PHA and PWM stimulated cells showed only minor quantitative differences when compared in silver stained gels, one qualitative and two quantitative differences between the two mitogens could be observed in fluorograms. Individual variability was generally low: within the approximately 30 unrelated individuals examined, thus far, the presence of two polymorphic proteins was demonstra

ISSN:0173-0835    

DOI:10.1002/elps.1150061207

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

摘要:

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 an

ISSN:0173-0835    

DOI:10.1002/elps.1150061208

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150061209

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150061210

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150061201

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1985

数据来源: WILEY