摘要:

AbstractHydrophobic affinity electrophoresis under high hydrostatic pressure has been developed to study the interaction between fatty acid‐free bovine serum albumin and a long‐chain aliphatic ligand physically immobilized within the gel matrix. From apparent association constants at various pressures and temperatures, apparent thermodynamic parameters including the volume change in binding were calculated. The results are as expected for hydrophobic interactions between the long‐chain alkyl ligand and a high‐affinity long‐chain fatty acid binding site. The feasibility of high‐pressure affinity electrophoresis is demonstrated. This new high‐pressure technique provides a direct means for studying quantitatively the effects of pressure upon protein‐ligand interactions. It could become a suitable tool for the investigation of protein binding si

ISSN:0173-0835    

DOI:10.1002/elps.1150090402

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

年代:1988

数据来源: WILEY

摘要:

AbstractCrude extracts of turnip crinkle virus upon agarose gel electrophoresis yield (i) virus patterns unperturbed by contaminants; (ii) plots of mobilityvs.gel concentration (Ferguson plots) parallel with those of the purified virus. The parallelism suggests similarity in size and shape but a lower net charge for the crude virus. This result is obtained when gel electrophoresis is carried out either in a continuous buffer or in a discontinuous (moving boundary electrophoresis) buffer system. The latter mode has the substantial benefit of electrophoretic (auto‐)concentration of dilute virus sample prior to resolution. Thus, the Ferguson plot analysis in a discontinuous buffer system of turnip crinkle virus can be viewed as a model procedure for the physical identification of other viruses contained in dilute extracts, feasible even in the absence of a prior knowledge as to the nature of, or isolation of, the viru

ISSN:0173-0835    

DOI:10.1002/elps.1150090403

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

年代:1988

数据来源: WILEY

摘要:

AbstractDiscontinuous buffer systems operative between pH 5.7 and 7.4, 0 °C, were generated, which are characterized by more rapidly displaced moving boundaries than applied previously. These allow one to resolve subcellular particles relatively rapidly and at relatively low agarose gel concentrations. A commercial mixture of DNA restriction fragments pre‐stained with ethidium bromide was found to be a suitable tracking dye for these boundari

ISSN:0173-0835    

DOI:10.1002/elps.1150090404

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

年代:1988

数据来源: WILEY

摘要:

AbstractImmobilized pH gradient gel rods, 1.5 mm in diameter, were cast with a manifold connected to high‐precision burettes. The reproducibility of gel length wasca.1.7 mm. The average standard deviation σxfor spot position was 2 mm after one‐dimensional and 5.8 mm after two‐dimensional runs. In order to bring to completion the elution of the salt fronts into the electrode compartments, carrier ampholytes had to be included in the gel formulation at concentrations of at least 0.5–1%, depending on the pH range. The presence of carrier ampholytes, however, was troublesome in two respects: the gel tended to shrink and the cathodic bands drifted with time. Ionic components in the sample were tolerated up to the following concentrations: NaCl 8 μmoles, sodium dodecyl sulfate 10 μg per tube. In presence of non‐ionic detergents, the gels moved as a whole towards

ISSN:0173-0835    

DOI:10.1002/elps.1150090405

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

年代:1988

数据来源: WILEY

摘要:

AbstractSerum was collected from rabbits at 2‐day intervals following a single injection with tetanus toxoid or at weekly intervals following multiple injections withMicrococcus lysodeikticuscell walls. These sera were analyzed for the presence of individual clonotypes of specific anti‐tetanus or anti‐micrococcal antibodies by isoelectric focusing in immobilized pH gradients with added carrier ampholytes followed by affinity immunoblotting. The affinity immunoblots obtained clearly defined both the rapid disappearance and late appearance of distinct subsets of antibody clonotypes during the response. These data demonstrate the application of affinity immunoblotting combined with immobilized pH gradients for detecting the subtle changes in specific antibody clonotype patterns which occur during an immune res

ISSN:0173-0835    

DOI:10.1002/elps.1150090406

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

年代:1988

数据来源: WILEY

摘要:

AbstractA base for a modern species' concept of chlorococcal algae can be obtained, not by morphological analysis, but by biochemical characters,e. g.isoenzyme banding patterns. From isolated lichen algae of the genusTrebouxiade Puymaly a set of five such enzymes has been studied by isoelectric focusing in immobilized pH gradients (IPG): phosphoglucomutase, phosphoglucose isomerase, malate dehydrogenase, mannitol dehydrogenase and leucine aminopeptidase. The first four are resolved into isoforms in a pH 4–7 IPG interval, while the last one is analyzed in an IPG pH 3.5–5 span. The patterns are specific for distinct populations, inter‐ and intraspecifically varying in dependence from their geographical distribution or the lichen species from which they have been isolated. Their limited heterogeneity (one to four isoforms) suggests that they are the products of specific genes rather than artefacts of the extraction procedure or the IPG analysis. Sharp isozyme patterns can only be obtained in a mixed‐bed, carrier‐ampholyte (CA)‐IPG gel and by anodic application, suggesting that the recently proposed mechanism of hydrophobic protein‐IPG matrix interaction (Electrophoresis, 1987,8, 62–70) is fully operative here. As an additional mechanism, it is proposed that, in some cases, CA might simply act, when added to an IPG gel, by buffering, in the transient state, the sample zone before the protein migrates from the liquid phase into

ISSN:0173-0835    

DOI:10.1002/elps.1150090407

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

年代:1988

数据来源: WILEY

摘要:

AbstractThe feasibility of detecting quantitative genetic variants based on a decrease in the integrated intensity of polypeptide spots in two‐dimensional polyacrylamide gels of human lymphoblastoid cell clones was investigated. A battery of 65 spots on 115 gels was studied to determine the distribution of quantitative measures for spots where no mutation had occurred. The corresponding distribution for spots which have decreased integrated intensity as a result of a mutation at one of two alleles coding for the spot was investigated by quantitating spots for which mutations were known to have occurred. These two distributions allowed the estimation of the rates of false positive and false negative errors for any particular strategy aimed at detecting null mutations, and thus provides a basis for the design of efficient strategies. Our silver stained gels have sufficient reproducibility of spot integrated intensities so that, for situations in which the mutation rate is relatively high, it is practical to monitor a sub‐set of spots for null variants using the same digitized images as are used to detect structural varia

ISSN:0173-0835    

DOI:10.1002/elps.1150090408

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

年代:1988

数据来源: WILEY

摘要:

AbstractWe recently established that every ion has two radii – a geometrical one and an electrokinetic one. Using this principle, and in accordance with Henry's method of determining his function, f(κa), we have discovered that the Henry function can be radically simplified. The graphs of the proposed equation are plotted at different argument values. They are identical with the graphs plotted according to the Henry functi

ISSN:0173-0835    

DOI:10.1002/elps.1150090409

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

年代:1988

数据来源: WILEY

摘要:

AbstractA quantitative theory of the Stern electric double layer is suggested. It is based on the view that every ion possesses a geometrical and an electrokinetic radius, that the ionic atmosphere begins from the geometrical one, and that the difference between these radii is the Stern quantity Δ. The equations of the mentioned radii and the quantity Δ are established and the values of the different potentials characterizing an ion and its ionic atmosphere are determine

ISSN:0173-0835    

DOI:10.1002/elps.1150090410

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.1150090411

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

年代:1988

数据来源: WILEY