ISSN:0173-0835    

DOI:10.1002/elps.1150090302

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

年代:1988

数据来源: WILEY

摘要:

AbstractImproved methods for high resolution composite gel electrophoresis under dissociating conditions and electrophoretic transfer of immunoglobulins (Mr150 000‐1 800 000) to nitrocellulose have been developed. Using these techniques and highly specific antisera to detect either light or heavy chains on nitrocellulose transfers, the immunoglobulin and clonal composition of washed cryoglobulins (3 μg each) could be determined within two days. The results were confirmed by isoelectric focusing and immunoblotting of selected samples. In addition, the method was used for detection of andiotypic determinant and rheumatoid factor activity within components of the cryoglobuli

ISSN:0173-0835    

DOI:10.1002/elps.1150090303

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

年代:1988

数据来源: WILEY

摘要:

AbstractWe previously analyzed unconcentrated cerebrospinal fluid (CSF) from patients with multiple sclerosis (MS) and other neurologic diseases by isoelectric focusing in agarose gel. We have now developed an immunoblot method for detection of oligoclonal IgG bands in unconcentrated MS CSF. The oligoclonal IgG band patterns seen after immunoblotting were compared with those of conventional immunofixation. Although immunoblotting was found to be rapid the resolution and intensity of oligoclonal IgG bands were somewhat better after immunofixation. Since immunofixation is simpler than immunoblotting, we recommend that clinical laboratories use immunofixation after isoelectric focusing to detect oligoclonal IgG bands in CSF.

ISSN:0173-0835    

DOI:10.1002/elps.1150090304

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

年代:1988

数据来源: WILEY

摘要:

AbstractHemolysates of normal, heterozygous hypocatalasemic and acatalasemic mice and of Japanese acatalasemic subjects were separated into three fractions, A, B and C, by DEAE‐cellulose column chromatography, and pIvalues of A, B and C fractions were determined by isoelectric focusing. The pIvalue of catalase in the A, B and C fractions increased in the order of normal, hypocatalasemic and acatalasemic mouse blood. The results obtained from Japanese acatalasemic blood samples showed that the pIvalues of catalase in the A, B and C fractions were similar to those in normal blood. Catalase in Japanese acatalasemic cultured skin fibroblasts was also analyzed by isoelectric focusing. The pIvalues of catalase in the extract from the cultured skin acatalasemic fibroblasts was similar to that in normal fibroblast

ISSN:0173-0835    

DOI:10.1002/elps.1150090305

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

年代:1988

数据来源: WILEY

摘要:

AbstractWe have resolved and characterized three forms of human and rat hepatic class III alcohol dehydrogenase. Separations were carried out in narrow immobilized pH gradients. Both in humans and rats the three forms were visualized by enzyme staining with cinnamol, but not with ethanol. They were insensitive to the inhibitory effect of pyrazole. The isoelectric points were approximately from 6.3–6.4, from 5.9–6.0 and 5.6. Each electroeluted enzyme extract, purified further by analytical isoelectric focusing over the pH range from 5–6 or 6–7, revealed a single band by enzyme and silver staining and by Western blotting followed by avidin‐biotin staining. Polyacryl‐amide gel electrophoresis in the presence of sodium dodecyl sulfate of each extract revealed a single molecular mass species corresponding to class III alcohol dehydrogenase (ADH). All forms of class III alcohol dehydrogenase were recognized by antisera raised against total cl

ISSN:0173-0835    

DOI:10.1002/elps.1150090306

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

年代:1988

数据来源: WILEY

摘要:

AbstractThe interpretation of two‐dimensional gel electrophoresis (2‐DGE) profiles can be facilitated by artificial intelligence and machine learning programs. We have incorporated into our 2‐DGE computer analysis system (termed MELANIE‐Medical Electrophoresis Analysis Interactive Expert system) a program which automatically classifies 2‐DGE patterns using heuristic clustering analysis. This program is a step toward machine learning. In this publication, we describe the classification method and the preliminary results obtained with liver biopsy electrophoretograms. Heuristic clustering is also compared to other classification t

ISSN:0173-0835    

DOI:10.1002/elps.1150090307

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

年代:1988

数据来源: WILEY

摘要:

AbstractThe proteins of cheese are rapidly solubilised by heating to 95 °C in buffered 2 % sodium dodecyl sulfate, 5 % 2‐mercaptoethanol. Electrophoretic analysis of the solubilised proteins by either one‐dimensional sodium dodecyl sulphate‐polyacrylamide gel electrophoresis or high resolution two‐dimensional electrophoresis yields reproducible patterns characteristic of an individual cheese and its extent of ripening. The patterns reveal (i) the residual amounts of milk casein and whey proteins, and (ii) the appearance of casein degradation products, including pink‐violet components as detected by Coomassie Blu

ISSN:0173-0835    

DOI:10.1002/elps.1150090308

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

年代:1988

数据来源: WILEY

摘要:

AbstractArtifacts associated with gold staining of polypeptides separated by sodium dodecyl sulphate – polyacrylamide gel electrophoresis and transferred to nitrocellulose are eliminated by alkylation of reduced samples or by deletion of the reducing agen

ISSN:0173-0835    

DOI:10.1002/elps.1150090309

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

年代:1988

数据来源: WILEY

摘要:

AbstractAn apparatus and the procedure for cellulose acetate isoelectric focusing were refined and optimized for routine use in clinical laboratories. The space in the electrophoretic chamber was minimized to maintain high humidity during the run. The water content of the thin cellulose acetate membranes in the chamber was well controlled during isoelectric focusing and the effect of atmospheric carbon dioxide was also excluded. The temperature of the membranes was kept below 2 °C, even under conditions of high electric field strength for faster isoelectric focusing of high‐molecular‐weight proteins. No special training was required for technicians in clinical laboratories to obtain reproducible isoelectric profiles of human serum prot

ISSN:0173-0835    

DOI:10.1002/elps.1150090310

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

年代:1988

数据来源: WILEY

摘要:

AbstractIn the course of a population study of α1‐antitrypsin polymorphism by separator isoelectric focusing, a variant phenotype having a somewhat narrower spacing than PI M1M3 was observed in a Japanese blood donor. Family studies by hybrid isoelectric focusing in a carrier ampholyte‐supplemented immobilized pH gradient from 4.35–4.65 revealed that the products of the responsible gene,PI*Mtoyoura, were extremely close but slightly cathodal to those ofPI*M1.The difference in isoelectric point between them corresponded to the resolving limit of isoelectric focusing. For this reason, although the propositus' father was deduced to have the genotypePI*M1/PI*Mtoyoura, the products of these two genes failed to form a double band pattern. Thus, the gene frequency forPI*Mtoyourawas unknown. These findings, however, indicate that a further microheterogeneity in thePI Msubtype exists at least in the Ja

ISSN:0173-0835    

DOI:10.1002/elps.1150090311

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

年代:1988

数据来源: WILEY