摘要:

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

ISSN:0173-0835    

DOI:10.1002/elps.1150060902

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

年代:1985

数据来源: WILEY

摘要:

AbstractNon‐ionic detergents (NP‐40, Triton X‐100) have poor solubilizing properties on membrane proteins, as demonstrated by separating NP‐40‐extracted proteins by isoelectric focusing also in the absence of carrier ampholytes (Ampholine),i. e.in immobilized pH gradients (IPG). Addition of 4% Ampholine to the sample and to the IPG gels results in an increase in membrane solubility, from 40% in NP‐40 alone to 60% in NP‐40 with 4% Ampholine, when both extractions were in the absence of 8 M urea. If the latter solvent is used, these values are increased to 85% for the former and to 100% for the latter extraction conditions. In presence of carrier ampholytes sharp bands are obtained on isoelectric focusing. It is hypothesized that neutral detergents in presence of Ampholine form mixed micelles, mimicking the properties of natural zwitterioni

ISSN:0173-0835    

DOI:10.1002/elps.1150060903

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

年代:1985

数据来源: WILEY

摘要:

AbstractThe phenotyping of the third component (C3) of human complement has been performed by isoelectric focusing in immobilized pH gradients followed by immunoblotting on nitrocellulose filter membrane. This powerful technique reveals variations of C3* and C3*F alleles not detected by agarose electrophoresis. The limits of the resolving power of isoelectric focusing in immobilized pH gradients for C3 analysis are shown to depend upon the high molecular weight of this protein. The notion of “suptypes” is discussed. Finally, the importance of subtyping for medical applications and for determination at the molecular level of interacting protein mechanisms is underli

ISSN:0173-0835    

DOI:10.1002/elps.1150060904

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

年代:1985

数据来源: WILEY

摘要:

AbstractA systematic analysis of protein staining in polyacrylamide gels with Coomassie Brilliant Blue (CBB) R‐250 and G‐250 using a high resolution densitometer allowing for quantitative measurements during staining and destaining has revealed that none of the published procedures allows quantitative measurements. Protein staining with CBB R‐250 in methanol/water/acetic acid is poor, as is staining with CBB G‐250 in trichloroacetic acid or perchloric acid, the latter two, however, allowing for a weak background staining. Consequently using the colloidal properties of the CBB dyes, stronger for G‐250 than for R‐250, it is possible to increase the sensitivity of protein staining to a detection limit of 0.7 ng bovine serum albumin/mm2gel. In addition, sensitive protein staining on a clear background is possible. Recipes are described (Section 3.11) for intensified protein staining with CBB G‐250 using trichloroacetic acid or perchloric acid on a clear background. Optimal staining of proteins on a clear background can be performed with phosphoric acid and CBB G‐250 in the presence of ammonium sulfate since under these conditions the colloidal state of the dye is optimized. Furthermore, conditions are described which allow the stable fixation of the protein‐dye complex. Combining the optimized staining conditions with the stable fixation in 20% ammonium sulfate allows for stepwise staining fore. g.detection of weak spots in addition to intense protein spots. The dependence of different staining procedures on gel thickness, gel concentration and compounds routinely used in polyacrylamide gel electrophoresis is also analysed. Calibration curves and application of the new procedure to biological material demonstrate its wide applicability. Convincing arguments for the colloidal properties of the CBB dyes are presented, formulating the rationale for intensified protein staining with CBB dyes in polyacrylamide gels without ba

ISSN:0173-0835    

DOI:10.1002/elps.1150060905

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

年代:1985

数据来源: WILEY

摘要:

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.

ISSN:0173-0835    

DOI:10.1002/elps.1150060906

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

年代:1985

数据来源: WILEY

摘要:

AbstractBuffer focusing using the mixture Polybuffer 47 (Polysciences) has been tested for the separation of proteins under denaturing conditions to assess its potential for use as the first dimension in two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE). a relatively simple pattern, with the proteins confined to certain regions of the gel, was observed when [35S]methionine‐labelled fibroblasts were separated both by 2D‐PAGE and by one‐dimensional isoelectric focusing (IEF) using Polybuffer 47. When Polybuffer 47 and synthetic carrier ampholytes were mixed, the latter were also found to concentrate into regions corresponding to those occupied by the proteins. The addition of synthetic carrier ampholytes resulted in separation of proteins within these regions. Diagonal IEF gels of synthetic carrier ampholyteversusbuffer focusing showed a “step” pattern, suggesting that buffer focusing resulted in concentration of proteins in particular regions. Diagonal IEF gels using synthetic carrier ampholytes in both dimensions verified that protein‐ampholyte interactions are not a major source of artefacts in such gels. When mixtures of Polybuffer 47 and synthetic carrier ampholytes were used, there was some extension of the pH gradient at both ends. We conclude that Polybuffer 47 alone is not suitable for use in the first dimension of high‐resolution 2D

ISSN:0173-0835    

DOI:10.1002/elps.1150060907

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

年代:1985

数据来源: WILEY

摘要:

AbstractHuman plasma proteins were subjected to parallel micro two‐dimensional polyacrylamide gel electrophoresis in the absence of denaturing agents. Forty nitrocellulose blots were obtained from eight slab gels within one hour of electroblotting time, employing an apparatus designed to obtain multiple replicas from micro slab gels. The locations of 31 human plasma proteins were determined on these blots using specific antisera, and an “identification map” was prepared. Differences of protein location between plasma and serum were also demonst

ISSN:0173-0835    

DOI:10.1002/elps.1150060908

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

年代:1985

数据来源: WILEY

摘要:

AbstractA routine method for species' determination of raw muscle tissues is described, Defrosted raw muscle tissues were evenly spread on filter paper applicators which were then placed on a thin‐layer agarose plate. A pH 4–8 gradient was obtained by mixing equal parts of Ampholine 4–6 and 6–8. Tissue homogenates of bovine, equine, porcine, ovine and other species were compared with spread tissue applications. The proposed method is useful in the routine control for quick identification of cut meat in cases of fraud where an exchange of species may have take

ISSN:0173-0835    

DOI:10.1002/elps.1150060909

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

年代:1985

数据来源: WILEY

摘要:

AbstractThe existence of a geometrical and an electrokinetical radius of an ion is suggested. An equation is worked out representing the electrokinetical radius as a function of the geometrical radius and the thickness of the ionic atmosphere.

ISSN:0173-0835    

DOI:10.1002/elps.1150060910

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

年代:1985

数据来源: WILEY

摘要:

AbstractA linear equation representing the ionic mobility as a function of the ionic strength square root was worked out on the basis of our hypothesis for the existence of two ionic radii. The equation contains a proposed parameter,pi. Comparisons between this equation and the equations of Onsager, and Robinson‐Stokes were made, and the absolute mobilities of 20 ions at 0 and 25 °C were calculated according to it. It was demonstrated that equations derived from the proposed equation can be used for calculating electrolyte dissociation degrees and electrolyte concentration dissociation constan

ISSN:0173-0835    

DOI:10.1002/elps.1150060911

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

年代:1985

数据来源: WILEY