摘要:

AbstractComputer‐aided quantitative analysis of densitometric spectra is presented. The densitometric spectra are decomposed into component bands using the Powell and Marquardt minimizers. Several different functions for the component bands are utilized. It is shown that the densitometric spectra can be decomposed into component bands with high accuracy only if the proper shapes of the bands are chosen. The method described was used for quantitative analysis of densitometric spectra of DNA cleaved by neocarzinostatin. The procedure is general and can be applied to analyses of autoradiographic films and to direct scans of electrophoretic gels. It is shown that densitometric spectra which show highly overlapped bands are well approximated by asymmetric Cauchy and Gauss functions. Well separated densitometric bands which have substantial asymmetry can be fitted to more sophisticated shapes formed by combinations of symmetric and asymmetric Cauchy and Gauss functions. High accuracy fitting to asymmetric densitometric curves may only be achieved using the cosine function introduced by Mignot and Rondot,J. Appl. Cryst.1976,9, 460–

ISSN:0173-0835    

DOI:10.1002/elps.1150101002

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

年代:1989

数据来源: WILEY

摘要:

AbstractNativeHemophilus influenzaepolysaccharide‐protein conjugate particles were analyzed by a two‐dimensional agarose electrophoresis procedure. In view of their preparation by random chemical crosslinking, the conjugates necessarily exhibit a polydisperse two‐dimensional gel pattern which varies depending on the conditions of the particular preparation. The polydisperse patterns were interpreted with regard to the size and surface net charge density of the conjugate on the basis of the extended Ogston model. Data processing was performed by a new program, designated ZWEIDI.DO, written in the language of M‐LAB (modeling laboratory). The program computes particle and gel fiber specific parameters from the positions of standards and unknown(s) on the two‐dimensional gel using a simultaneous linear least‐square curve fitting routine. Based on these calculations, the program serves to compute a nomogram of iso‐size and iso‐free‐mobility profiles. Superimposing these profiles on the gel patterns, the size and free mobility range of the polydisperse conjugate mixtures is obtained. Potentially, the procedure could serve as a tool for quality control in the production of conjugates as vaccines and for the physical characterization of polydisperse subcellular parti

ISSN:0173-0835    

DOI:10.1002/elps.1150101003

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

年代:1989

数据来源: WILEY

摘要:

AbstractA computer‐assisted statistical treatment of the electrophoretic data obtained from the analysis of two dehydrogenases and 27 kinds of esterases produced by strains belonging to the taxonomically complex genusAcinetobacteris described. The 12 genospecies were clearly separated from each other by correspondence analysis, For each genospecies the distances of the strains from their barycenter were computed and typical isolates suitable for use as reference strains were determined. This approach is suitable for the systematic study of other procaryotic or eucaryotic organism

ISSN:0173-0835    

DOI:10.1002/elps.1150101004

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

年代:1989

数据来源: WILEY

摘要:

AbstractA theory for discontinuous electrophoresis in a polyacrylamide gel is presented for one buffer at two pH values. It is shown that polyions stack between identical leading and trailing ions, and resolve in a gel of constant polyacrylamide concentration. The theory is illustrated by the separation of serum protein polyions in a Tris‐glycinate buffer of pH 8.19 in the well‐forming gel, and pH 9.16 in the resolving gel. The selected concentrations and electrolyte ionization degrees of Tris and glycine have values at which the serum protein polyions stack between the resolving and electrode buffers, followed by separation in the resolving

ISSN:0173-0835    

DOI:10.1002/elps.1150101005

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

年代:1989

数据来源: WILEY

摘要:

AbstractIsolated metaphase chromosomes of several fibroblastoid cell lines (Chinese hamster, Chinese hamster × human hybrid) were subjected to free flow electrophoresis (FFE) to study their electrophoretic mobility (EM). The morphology and stability of the chromosomes were unaffected by FFE as examined by cytogenetic methods and flow cytometry. The chromosomes of the complement all showed similar EM under most of the conditions applied. At neutral pH the EM of the chromosomes had the same sign as free DNA and about 2/3 of its magnitude. The variation of EM with buffer parameters such as ionic strength, valence of counterions, buffer capacity and dielectric constant of the solvent were investigated. Thermal denaturation increased the EM of the chromosomes by 20 %. Partial denaturation might offer a possibility to separate or enrich large amounts of chromosomes by FFE

ISSN:0173-0835    

DOI:10.1002/elps.1150101006

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

年代:1989

数据来源: WILEY

摘要:

AbstractThe impact of the type of membrane used to separate the electrolyte compartments from the separation cell for continuous flow field step electrophoresis is examined using both experimental and computer simulation data. It is shown that the distinct transport characteristics of ion exchange membranes and dialysis membranes may have an effect on the stability of the pH and conductivity gradients created by the buffer system. These effects are interpreted using computer simulation data. The limitations of the model used to portray the transport characteristics of ion exchange membranes are discussed as are the experimental conditions which provide the greatest stability for field step electrophoresis.

ISSN:0173-0835    

DOI:10.1002/elps.1150101007

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

年代:1989

数据来源: WILEY

摘要:

AbstractThe electrophoretic titration curves of complex mixtures of vitamin K‐dependent human blood proteins and proteins ofBothrops aspervenom were investigated. In both protein mixtures some curves exhibited marked distortions such as additional maxima and minima when Pharmalyte 3–10 carrier ampholytes were used for isoelectric focusing in agarose gels. The distortions result from an unspecific interactions‐between some carrier ampholyte constituents with particular proteins. The interacting carrier ampholyte components could be completely removed by binding to albumin and ultrafiltration through a UM‐2 Amicon membrane with resultant regular titration curves. The interacting carrier ampholyte species were only partially removed by ultrafiltration through a UM‐2 membrane without incubation wit

ISSN:0173-0835    

DOI:10.1002/elps.1150101008

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

年代:1989

数据来源: WILEY

摘要:

AbstractThe methodology for conventional radiofluorography of two‐dimensional gels, followed by rehydration of the gel and subsequent silver staining, is described. The image obtained by radiofluorography is referred to as biosynthetic image, and the image obtained by silver staining as constitutive image. Since the two images are already in close register (the same gel), reliable identification of polypeptides by the two different assays is possible, and the comparison provides valuable information on the catabolism of each entity. The utility of this procedure is illustrated in experiments involving a labeling with L‐[35S] methionine of an entire mouse. Both serum and tissue samples were analyzed by two‐dimensional gel electrophoresis with the aim of determining several categories of polypeptides in terms of their biosynthetic rates and their compos

ISSN:0173-0835    

DOI:10.1002/elps.1150101009

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

年代:1989

数据来源: WILEY

摘要:

AbstractA multiple immunoblotting technique was developed to positively identify up to three different antigens on a single nitrocellulose replica of a two‐dimensional isoelectric focusing/sodium dodecyl sulfate‐polyacrylamide gel. Three highly sensitive immunoblot assays were selected, including: horseradish peroxidase/luminescence, alkaline phosphatase, and silver‐enhanced immunogold. As a major advantage, the method permits a simultaneous detection of up to three different antigens without eluting the antibody‐dye complex between staining of single polypeptides, thus providing a highly accurate identification of closely migrating components. The staining procedure is summarized in a flow chart. In addition to the multiple immunoblot staining, some suggestions are provided for a sensitive protein s

ISSN:0173-0835    

DOI:10.1002/elps.1150101010

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

年代:1989

数据来源: WILEY

摘要:

AbstractNonequilibrium agarose isoelectric focusing in a pH 4.5–7 gradient with 3‐(N‐morpholinopropanesulfonic) acid (MOPS) and taurine as chemical separators presents a fast, easy and reliable method for the simultaneous determination of the common erythrocyte acid phosphatase and esterase D phenotypes in hemolysates and dried bloodstains. The application of the method to experimental and casework bloodstains is desc

ISSN:0173-0835    

DOI:10.1002/elps.1150101011

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

年代:1989

数据来源: WILEY