摘要:

AbstractIn crossed immunoaffinoelectrophoresis with free concanavalin A in the first dimension, human serum α1‐antichymotrypsin, purified or in whole serum, exhibited four peaks in presence of 0.02 M α‐methyl‐D‐glucoside added to the second‐dimensional gel. α1‐Antichymotrypsin purified from the serum of a single healthy donor was separated by affinity chromatography into three fractions on a laboratory‐prepared concanavalin A‐Sepharose 4B column: a pass‐through fraction, a retarded fraction and a bound fraction, eluted from the column on addition of sugar to the buffer. These three fractions were analyzed by crossed immunoaffinoelectrophoresis and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and isoelectric focusing before and after desialylation. The results of electrophoresis as well as chemical analyses indicate that these microheterogeneous forms carry glycans with decreasing degrees of branching from the concanavalin A‐pass‐through form to the concanavalin A‐bound form. This approach represents a first step towards the elucidation of the molecular basis of the microheteroge

ISSN:0173-0835    

DOI:10.1002/elps.1150100402

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

年代:1989

数据来源: WILEY

摘要:

AbstractR‐Form lipopolysaccharides ofAcinetobacter calcoaceticuscould be incorporated into polyacrylamide gels in an immobile form by adding it directly to the acrylamide‐N, N′‐methylenebisacrylamide polymerization mixture. The separation ofA. calcoaceticus69 V outer membrane proteins in these affinity gels demonstrated a specific interaction with the lipopolysaccharide ligand for one of the proteins. This protein is heat‐modifiable and has anMrof about 18 000. By incorporation of varying concentrations of lipopolysaccharide, a dissociation constant of the protein‐lipopolysaccharide complex of 0.5 mM could be determined. In comparison, for anotherA. calcoaceticusstrain, CCM 5593, a higher dissociation constant (1.0 mM) – indicative of lower affinity

ISSN:0173-0835    

DOI:10.1002/elps.1150100403

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

年代:1989

数据来源: WILEY

摘要:

AbstractThe influence of a soluble anionic polymer on electrophoresis of proteins was studied in relation to the nonspecific ionic effect of an affinophore on application to affinophoresis. Zone electrophoresis of proteins was carried out in agarose gel in the presence of succinyl‐poly‐L‐lysine (degree of polymerization, 120) by using three electrophoresis buffers differing in ionic strength (0.06, 0.12 and 0.18) and pH (7.0 and 7.9). Proteins migrated as distinct single bands even in the presence of the polymer. The mobility of cationic proteins towards the cathode was first decreased and then increased towards the anode as the polymer concentration increased, while that of anionic proteins was not affected. The dependence of the apparent mobility changes of the proteins on the concentration of the polymer was treated quantitatively in the same way as affinity electrophoresis. The extent of the ionic interaction between a cationic protein and the polymer could be estimated as an apparent dissociation constant. It greatly depended on the ionic strength of the electrophoresis buffer. Except for the extremely cationic proteins such as lysozyme, the ionic interaction with up to 0.1 mM of the polymer could be practically suppressed by the use of 0.1 M sodium phosphate buffer (pH

ISSN:0173-0835    

DOI:10.1002/elps.1150100404

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

年代:1989

数据来源: WILEY

摘要:

AbstractThe effect of blocking nitrocellulose membranes with the nonionic detergent Tween 20 on the detection, by protein blotting, of IgE‐binding to house dust miteDermatophagoides pteronyssinusallergens has been investigated. Tween blocking led to enhanced immunodetection of allergens despite removal of proteins from the membrane when compared to protein blocking agents which did not displace transferred components. The enhancement varied with the different mite components and, for one in particular, antigenDer pII, an increase of more than 100‐fold in IgE antibody binding occurred despite a concurrent loss of more than 90% ofDer pII from the membrane. Both the enhancement of binding and loss of components from the membrane were dependent upon the time course of blocking and the concentration of Tween u

ISSN:0173-0835    

DOI:10.1002/elps.1150100405

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

年代:1989

数据来源: WILEY

摘要:

AbstractAn easy‐to‐perform and efficient blotting method was developed for the transfer of proteins separated in PhastGel media. The method was evaluated by comparing the blotting results of low molecular weight marker proteins at different times.14C‐Labelled proteins were used for the assessment of the transfer efficiency. Highly efficient transfer was achieved within 1 h of blotting and the proteins were almost completely eluted from the gel in 2 h. The gel remained on its solid plastic backing, which is not the case when using an electrophoretic transfer method. As little as 1.25 ng of electrophoretically separated protein were enzymatically detected on the blot when applying blot‐biotinylation. The transfer was performed at low temperature (4°C), which – in contrast to thermoblotting – additionally offers the chance to blot thermolabile proteins. These superior features, together with the method's versatility and low cost, mark an interesting alternative to the previously recommended blotting procedures after PhastSystem ele

ISSN:0173-0835    

DOI:10.1002/elps.1150100406

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

年代:1989

数据来源: WILEY

摘要:

AbstractThe representative β‐hydroxyethylmorpholinium‐chloride‐bicinate moving boundary with a trailing ion net mobility relative to Na+of 0.41, detected by precipitation of chloride with silver nitrate, exhibits a decreasing chloride mobility at increasing polyacrylamide gel concentrations from 3.5 to 45 %T, 5 %CBis. This decrease, largely due to an increase of field strength at constant current, is described by a convex plot of log (mobility)vs.%T (Ferguson plot) and signifies that chloride/bicinate are sieved by the gel. In agarose gels, the same plot of mobilityvs.gel concentration is constant below 7 % gel concentration, since in those gels field strength and migration rate remain the same within that gel concentration range. Both in polyacrylamide and in agarose gels the displacement rate of the chloride‐bicinate boundary as a function of the time of electrophoresis or distance migrated remains invariant within 15 %. The plot of log(mobility)vs.gel concentration extrapolated to 0 %T is 5.85 and 5.41 (10−5cm2s−1V−1) for polyacrylamide and for agarose (SeaKem HGT‐P, FMC) gels, respectively. The slightly decreased mobility intercept at 0 %T for agarose is presumably due either to the electroendosmotic properties of agarose HGT‐P and/or failure to Sufficiently take into account the flattening of the Ferguson plot in the polyacrylamide concentration range below 3 % in which a transition from a gel to a fluid (sol)

ISSN:0173-0835    

DOI:10.1002/elps.1150100407

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

年代:1989

数据来源: WILEY

摘要:

AbstractSodium dodecyl sulphate‐polyacrylamide gel electrophoresis has been used to determine the metacercariae (Mc) total body protein band profiles of differentDiplostomumspp. Four species of fish were investigated, roach(Rutilus rutilus)infected withD. spathaceumMc, gywniad(Coregonus laveratus)infected withD. coregonusMc, ruff(Gymnocephalus cernua)infected with aDiplostomumspecies related toD. gasterosteiMc, and perch(Perca fluviatilis)infected withD. gasterosteiMc. The four species ofDiplostomumMc were distinguished by three different bands of molecular weight,Mr55 500, 53 500 and 52 000. A homology of polypeptide component distribution was evident forD. gasterosteiMc, fromP. fluviatilis, andD. coregonusMc, fromC. laveratus, the latter showing reduced protein concentrations between bandsMr32 000–40 000. Critical analysis of Mc polypeptide patterns showed no evidence of contamination with lens, retina or vitreous humour host eye protein. Gross morphological data for the parasites was also considered in relation to different band profiles obtained for theDiplostomum spp.Mc. Band profile analysis in conjunction with other taxonomic tests proved to be a useful tool in the identification of unknown spec

ISSN:0173-0835    

DOI:10.1002/elps.1150100408

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

年代:1989

数据来源: WILEY

摘要:

AbstractAn algorithm for automatic evaluation of nucleic acid sequencing gel antoradiographs is described which is simple and fast, with a low level error rate.

ISSN:0173-0835    

DOI:10.1002/elps.1150100409

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

年代:1989

数据来源: WILEY

摘要:

AbstractA method was developed for the preparation of completely intact plant DNA embedded in agarose, and suitable for restriction enzyme digestion. Digestion with restriction enzyme was carried out according to modified protocols of Anand [1] and Kenwricket al. [2]. The new method of DNA isolation allows the separation of high molecular weight plant DNA by pulsed field gel electrophoresis.

ISSN:0173-0835    

DOI:10.1002/elps.1150100410

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

年代:1989

数据来源: WILEY

摘要:

AbstractHuman saliva was analyzed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and the response to different natural stimuli investigated. Serva Blue R revealed pink/violet‐staining proteins (“lumicarmines”). These intensified following stimulation but in an unpredictable and sometimes disproportionate manner, causing pattern changes. This variable response undermines their potential forensic value for suspect identif

ISSN:0173-0835    

DOI:10.1002/elps.1150100411

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

年代:1989

数据来源: WILEY