摘要:

AbstractAccurately standardized commercial polystyrene sulfate particles in agarose gel electrophoresis yield linear Ferguson plots at pH 7.4 over a gel concentration range up to 0.9% agarose which do not exhibit any significant sigmoidal curve elements, using either a discontinuous buffer system or a continuous buffer. Ferguson plots of these standard‐sized particles were evaluated using alternatively a linear or convex model, by means of a newly developed set of programs (to be used in conjunction with program M‐LAB) which (i) is sufficiently user‐friendly to allow for quantitative agarose gel electrophoresis of subcellular‐sized spherical particles based on their convex Ferguson plots with the same operational simplicity previously available for linear Ferguson plots only; (ii) simultaneously and interactively analyzes the Ferguson plots of all particles under consideration on the basis of an extended Ogsto

ISSN:0173-0835    

DOI:10.1002/elps.1150090602

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

年代:1988

数据来源: WILEY

摘要:

AbstractAn improved procedure for staining of proteins following separation in polyacrylamide gels is described which utilizes the colloidal properties of Coomassie Brilliant Blue G‐250 and R‐250. The new method is based on addition of 20 % v/v methanol and higher concentrations of ammonium sulfate to the staining solution previously described [1]. The method combines the advantage of much shorter staining time with high sensitivity, a clear background not requiring destaining, stepwise staining, and stable fixation after staining. The method has been applied to staining of polyacrylamide gels after sodium dodecyl sulfate‐electrophoresis and isoelectric focusing in carrier ampholyte‐generated pH gr

ISSN:0173-0835    

DOI:10.1002/elps.1150090603

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

年代:1988

数据来源: WILEY

摘要:

AbstractA simple method of flattening and/or expanding of pH gradients in isoelectric focusing is described for any pH interval desired: to modify pH gradients near one electrode a paper strip soaked with carrier ampholytes is applied onto the gel close to the opposite electrode. In order to flatten central parts of pH intervals paper strips are applied onto the gel at both electrodes. Conditions and criteria (e.g.amount and pH intervals of carrier ampholytes, width and localization of the paper strip, separation period) for optimization are presented with PhastSystem using ready‐made gels with three different pH intervals and pImarker proteins (Pharmacia). Examples utilizing erythrocyte lysates are presente

ISSN:0173-0835    

DOI:10.1002/elps.1150090604

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

年代:1988

数据来源: WILEY

摘要:

AbstractOrosomucoid (ORM) polymorphism was investigated by different methods including isoelectric focusing in acid pH ranges followed by silver staining, print immunofixation of desialyzed ORM, fixation using a lectin from the sea‐weedCodium tomentosum, isoelectric focusing followed by immunofixation in miniaturized gels and isoelectric focusing in immobilized pH gradients. Population genetics studies were carried out in Galicia (NW Spain) and two new ORM variants were foun

ISSN:0173-0835    

DOI:10.1002/elps.1150090605

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

年代:1988

数据来源: WILEY

摘要:

AbstractCrossed immunoelectrophoresis of human serum revealed two heterogeneity types of ceruloplasmin with different electrophoretic migration. The two types both consisted of peptides withMr150 000, 100 000 and 45 000, which were interpreted as native ceruloplasmin and two hydrolytic fragments. The two types were different in copper content, and one type could reversibly be changed into the other. The glycan microheterogeneity of ceruloplasmin was analyzed by crossed affinommunoelectrophoresis with freeLens culinarisagglutinin (LCA) and wheat germ agglutinin (WGA). A third of the ceruloplasmin molecules, both high and low copper type, bound to LCA and two thirds to WGA. The heterogeneity and the microheterogeneity of ceruloplasmin in two groups of patient sera were compared to sera from healthy individuals. The ceruloplasmin type with respect to copper content was a much better factor than either glycan microheterogeneity or total serum concentration in discriminating between the three groups.

ISSN:0173-0835    

DOI:10.1002/elps.1150090606

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

年代:1988

数据来源: WILEY

摘要:

AbstractA previously described double‐label two‐dimensional electrophoresis procedure (Wheeleret al., Anal. Biochem.1986159, 1–7) for the analysis of differences between two complex mixtures of soluble proteins has been modified to allow analysis of proteins requiring detergent for aqueous solubility. The samples are first disrupted by sonication and the insoluble proteins concentrated by high‐speed centrifugation. The proteins are then solubilized with sodium dodecyl sulfate and further concentrated in a centrifugal concentrator to achieve protein mixtures suitable for labeling with14C and3H by reductive methylation and subsequent two‐dimensional electrophoresis. The sample concentration step is quick, minimizes the concentration of sodium dodecyl sulfate in the final sample, and avoids the potential difficulties associated with lyophilization or precipitation. The modified procedure was applied to the analysis of erythrocyte membranes, platelets and isolated placental microvilli. The high resolving power of two‐dimensional gel electrophoresis is retained and the procedure is sensitive because the conditions of labeling allow substantial incorporation of radioactivity into protein despite the presence o

ISSN:0173-0835    

DOI:10.1002/elps.1150090607

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

年代:1988

数据来源: WILEY

摘要:

AbstractHigh sensitivity and low background, the attractive characteristics of the procedure of Blumet al.,Electrophoresis1987,8, 93–99, for silver staining of proteins in polyacrylamide gels have been improved by sensitizing the gels with sodium dithionite instead of sodium thiosulfate and by equilibration in water after fixation and prior to sensitization. These modifications decrease the background and allow for a longer development period, which in turn increases sensitivity and color contrast. In addition, the colors of the spots are shifted toward colder tones when compared with the original metho

ISSN:0173-0835    

DOI:10.1002/elps.1150090608

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

年代:1988

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150090609

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

年代:1988

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150090610

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

年代:1988

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150090601

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

年代:1988

数据来源: WILEY