ISSN:0173-0835    

DOI:10.1002/elps.1150060102

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

年代:1985

数据来源: WILEY

摘要:

AbstractA free fluid zone electrophoresis experiment was performed in the microgravity environment of Space Shuttle flight STS‐3 (March 1983). The experiment was designed to confirm observations made on the Apollo‐Soyuz mission of 1975 and to test the effect of high red blood cell (RBC) concentration on free fluid electrophoresis. Photographic documentation of cell zone progression in one‐hour separations of mixtures of formaldehyde‐fixed human and rabbit erythrocytes (RBC), which were subjected to a field of approximately 13 V cm−1in low ionic strength buffer, was analyzed. One of two columns contained 2 × 108RBC ml−1(low concentration), and the other contained 1 × 109RBC ml−1(high concentration). The observed and calculated leading edge displacements of the RBC in the two columns were in agreement, indicating the absence of unexpected effects of the reduced gravity environment. Post‐flight analyses of the contents of the columns was not possible, and additional microgravity experiments are needed to evaluate the role of particle‐particle interactions in concentrated suspensions undergo

ISSN:0173-0835    

DOI:10.1002/elps.1150060103

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

年代:1985

数据来源: WILEY

摘要:

AbstractThis paper deals with electrolyte systems capable of producing suitable isotachophoretic zone structures in either migration direction. These systems contain moving reaction boundaries. A dilute solution of a strong acid as anolyte and the sodium salt of a weak acid as catholyte are analyzed to provide insight into the features of such dual systems. For various electrolyte arrangements the boundaries are examined for resolving capability in isotachophoresis. Both anionic and cationic boundaries were followed simultaneously using a capillary type apparatus with an array detector.

ISSN:0173-0835    

DOI:10.1002/elps.1150060104

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

年代:1985

数据来源: WILEY

摘要:

AbstractA modified method of ultrathin‐layer isoelectric focusing is described. The method allows subtyping of phosphoglucomutase even in samples containing considerable amounts of salts and proteins without pretreatment. Wedge‐shaped polyacrylamide or agarose gels varying in thickness from 50 μm (cathode) to 300 μm (anode) are used. If the right pH gradient is chosen the isoenzymes will focus near the cathode where the gel is ultrathin. The sample is applied anodally where the gel is thickest and the capacity therefore high. Thus the effect of salts and proteins on the pH gradient is diminished. Comparative tests have shown that this method is superior to the conventional techniques using gels with a uniform thickness of 1

ISSN:0173-0835    

DOI:10.1002/elps.1150060105

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

年代:1985

数据来源: WILEY

摘要:

AbstractEnhanced separation of phosphoglucomutase (PGM1) isoenzymes has been achieved by addition of the separator N‐(2‐Hydroxyethyl)piperazine‐N′‐3‐propanesulphonic acid (EPPS) to pH 5–7 Ampholines in ultrathin and 1 mm isoelectric focusing gels. The resultant pH gradient ranges between pH 5.6 to pH 5.9 and effectively spreads PGM1isoenzymes across the whole width of the gel. The technique has been shown to be especially useful for separating some of the rare PGM1variants. The use of different electrolyte combinations has also been investigated and it has been shown that strong electrolytes,e. g.sodium hydroxide and orthophosphoric acid, result in a small but statistically significant increase in the gradient range compared with weaker electrolytes such as threonine/histidine. In addition, the effect of temperature on the rate of migration of PGM1isoenzymes has been

ISSN:0173-0835    

DOI:10.1002/elps.1150060106

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

年代:1985

数据来源: WILEY

摘要:

AbstractA silver stain method has been developed for isoelectric focusing in agarose gel. Its sensitivity is shown to be greater than 20 times that of Coomassie Brilliant Blue G‐250. Its usefulness in analyzing unconcentrated cerebrospinal fluid is demonstrate

ISSN:0173-0835    

DOI:10.1002/elps.1150060107

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

年代:1985

数据来源: WILEY

摘要:

AbstractA procedure for the electroimmunoassay of sodium dodecyl sulfate (SDS)‐solubilized proteins is described. The analysis of the small subunit of ribulose 1,5‐bisphosphate carboxylase in antibody‐containing gels was best achieved in presence of low SDS concentration. Polyethylene glycol, low electroendosmosis agarose and medium voltage were required for optimized assay. Bovine serum albumin can also be analyzed in SDS/antibody gels. The results suggest that the electroimmunoassay in SDS‐containing gels could be used for many p

ISSN:0173-0835    

DOI:10.1002/elps.1150060108

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

年代:1985

数据来源: WILEY

摘要:

AbstractGlycosaminoglycans, separated electrophoretically and stained with Alcian Blue 8GX, were quantified by densitometry. The method allows quantitation of all classes of glycosaminoglycans in samples containing up to 10 mg/ml of serum proteins. Detection sensitivity is 40–300 ng and the optimum range of analysis is either 40–400 or 300–2100 ng, depending on the classes of glycosaminoglycans examined. The method is based on the densitometric comparison between a reference standard of 150 ng of hyaluronate and biological samples electrophoresed on the same sheet. By densitometry the total glycosaminoglycan concentration as well as the percentage and concentration of single fractions are determined. Each class of glycosaminoglycans presents different affinity to Alcian Blue, resulting in over‐ or underestimation in comparison with hyaluronate. Such error is constant over the whole range of determination and may be corrected by correction factors evaluated for each class. Both electrophoresis and densitometric quantitation may be optimized for an analysis of different glycosaminoglycans extracted from biological samples. For instance, heparin and heparan sulfate may be identified by nitrosation applied directly on the sheet; each class may be quantified by comparison with its own standard glycosaminoglycan, avoiding over‐ or underestimation; moreover, dermatan sulfate may be subdivided into fractions. The method is suitable for the quantitation of glycosaminoglycans present in low concentration in biologica

ISSN:0173-0835    

DOI:10.1002/elps.1150060109

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

年代:1985

数据来源: WILEY

摘要:

AbstractThe polypeptide patterns obtained from individual caryopses (kernels) of two wheat cultivars were analyzed using high resolution two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE) methods. While few polypeptides were detected with Coomassie Blue staining methods, over two hundred peptides were easily distinguished with the color‐based silver stain. The relative isoelectric points of the constituent polypeptides ranged from pH 3.8 to 9 and the relative molecular weights from 5000 to 200 000. The majority of the proteins of the two wheat varieties analyzed with 2‐D PAGE were similar although distinct proteins having molecular weights of 23 000, 30 000, 37 000 and 70 000 were identified, which have molecular properties unique to each variety. The use of the color‐based silver stain makes it possible to identify and characterize hundreds of proteins in individual wheat seeds. These methods are, therefore, adaptable for the rapid analysis and characterization of specific gene products of single kernels

ISSN:0173-0835    

DOI:10.1002/elps.1150060110

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

年代:1985

数据来源: WILEY

摘要:

AbstractA stand is described which supports racks of gels for isoelectric focusing (ISO‐gels) and makes it possible to load these gel rods directly onto the second dimensional slabs of polyacrylamide (DALT‐gels). Equilibration with second dimensional buffer is made by adding it without agarose to the ISO‐gels on top of the slabs. A new design of DALT‐tank is also described to run twenty slabs of polyacrylamide while holding the plates vertically so the samples run down the slab. This system removes the need to seal the ISO‐gels onto the slab of polyacrylamide using agarose solutions. The omission of agarose reduces the background when two‐dimensional gels are stained

ISSN:0173-0835    

DOI:10.1002/elps.1150060111

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

年代:1985

数据来源: WILEY