摘要:

AbstractIntroduction of a linear of detectors along the electrophoretic separation space permits monitoring the evolution of electrophoretic zone structures, fully controlled by a microprocessor system. Both transient and steady state zone structures can be recognized. A description of our capillary type apparatus, with 255 potential gradient sensors measuring the most general physical property characterizing the separand pattern in electrophoresis, is given. The central unit of this device is a unique pattern of 256 electrodes. They are chemically photoetched on a supporting glass plate from a vapor‐deposited thin layer of a conductor. Each sensor has a width of 60 μm, a height of about 0.1μm and they are spaced 340 μm apart. The whole separation cell, appropriate detection electronics and a new constant current high tension power supply are described together with the microcomputer system used for control and data treatment. Illustrations of detected zone structures with steady state moving boundaries are given in terms of isotachophoretic and moving boundary experiments. The monitoring of non‐steady state boundaries is shown by following sample zones in zone electroph

ISSN:0173-0835    

DOI:10.1002/elps.1150050602

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

年代:1984

数据来源: WILEY

摘要:

AbstractA theory is described which gives a new algorithm for the selection of suitable electrolyte systems based on assuming (i) the zone stability as its correct isotachophoretic migration; (ii) the separability as the establishment of a fixed migration order of the zones of individual substances. The theory is based on data of the typeūi,j,i. e., effective mobility of substanceiin thej‐th zone. For checking the stability of zones, the use of the zone existence diagrams is demonstrated, where the groups of substances giving stable zones are easily characterized, with special attention to anionic analyses with OH−as the terminator. For checking the separability of substances, the use of the sequence contours in an existence diagram is demonstrated with respect to the zone order as well as to the formation of stable mixed zones. Experiments are given where anionic isotachophoresis of some amino acids with OH−as the terminator is shown, and the formation of stable mixed zones of substances with mutually different effective mobilities is demons

ISSN:0173-0835    

DOI:10.1002/elps.1150050603

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

年代:1984

数据来源: WILEY

摘要:

AbstractGel electrofocusing (EF) on Sephadex G‐75, which provides relatively non‐restrictive pore sizes and relatively broad protein concentration distributions (which reduces concentration‐dependent aggregation), was adapted for use in glass tubes. This allows for EF with meticulously controlled low temperatures in conjunction with large sample and gel volumes. Specifically, the technique was developed for EF of steroid receptors, which are highly sensitive to temperature (temperature transformed), and whose aggregation behavior excluded the use of polyacrylamide or agarose gels. The technique uses the temperature‐controlled apparatus designed for quantitative polyacrylamide gel electrophoresis, with three additions. These are a) a 12‐place evacuation device for forming simultaneously 12 cylindrical Sephadex gels (either 6 or 18 mm diameter), b) an electrode arrangement capable of maintaining the same regulated voltage across several gel tubes, and c) a jacket and lid of polyurethane foam to insulate the electrolyte chambers of the EF

ISSN:0173-0835    

DOI:10.1002/elps.1150050604

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

年代:1984

数据来源: WILEY

摘要:

AbstractSilver staining was examined as a means for detecting antigenic polypeptides separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) after recovery from immunoprecipitates produced by crossed immunoelectrophoresis and liquid precipitation. Both precipitin procedures resulted in detection of the expected polypeptides with bovine serum albumin (BSA) and human transferrin as model antigens, as well as microheterogeneity of BSA, whereas antigenic polypeptides were rarely detected with Coomassie Brilliant Blue staining of SDS‐PAGE. The technique is advantageous over the use of radiolabeled antigens in being less expensive and time consuming and over antibody‐blot procedures in detecting antigenic activity prior to the denaturing conditions of SDS‐PAGE. Possible drawbacks are the appearance of antibody polypeptide chains on the electrophorograms as well as other non‐antigenic components when crude extracts of virally‐infected cells are used

ISSN:0173-0835    

DOI:10.1002/elps.1150050605

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

年代:1984

数据来源: WILEY

摘要:

AbstractWe describe the use of35S‐labelled stains for the detection of submicrogram amounts of protein in SDS polyacrylamide gels. Many stains used to detect protein can be rendered radioactive, and proteins can be detected by radiofluorography due to stain binding in the absence of visible color. Of the stains tested. [35S]Ponceau Red S and [35S]Serva Blue R gave the best results. The potential utility of protein detection with radioactive stains is discusse

ISSN:0173-0835    

DOI:10.1002/elps.1150050606

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

年代:1984

数据来源: WILEY

摘要:

AbstractDansylated proteins have been used in combination as isoelectric point and molecular weight standards and included in gel samples for separation by two‐dimensional polyacrylamide gel electrophoresis. These proteins provide invariant points of reference that facilitate the unequivocal identification of proteins separated by two dimensional polyacrylamide gel electrophoresis without affecting resolution. Moreover, immunoautoradiograms of Western blots can be aligned with their corresponding gel patterns with great accuracy. The logical extension of the established usage of dansylated protein standards to these applications is describe

ISSN:0173-0835    

DOI:10.1002/elps.1150050607

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

年代:1984

数据来源: WILEY

摘要:

AbstractHigh resolution two‐dimensional polyacrylamide gel electrophoresis has been used to detect a soluble protein in extracts of human kidney which was not present in the electrophoretograms of other human tissues. This protein, coded kidney‐acidic protein‐60 (KAP‐60) was purified to homogeneity and was characterized as having a native molecular weight of 85000, a monomer molecular weight of 35 000 and an isoelectric point of 5.6. KAP‐60 was devoid of detectable carbohydrate or phosphate groups. Injection of the protein into rabbits produced antisera which were used to develop a radioimmunoassay for the protein and to localize the protein to specific cells within the kidney by the immunoperoxidase technique. A survey of human tissues by radioimmunoassay showed that KAP‐60 was present in human kidney at approximately 150 times the concentration found in human liver and 300 times the level found in other human organs examined. Immunohistochemistry localized the protein to the proximal and collecting tubules in the kidney with no staining of glomeruli or distal tubules. The present study illustrates the use of high‐resolution two‐dimensional electrophoretic techniques in the detection of human tissue‐specific proteins which may prove of clinical value in the biochemical diagnosis of tissu

ISSN:0173-0835    

DOI:10.1002/elps.1150050608

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

年代:1984

数据来源: WILEY

摘要:

AbstractA technique for the preparation of ultrathin titration curve gels, 50 μm polyacrylamide gels and 100 μm agarose gels, is presented. By separating protein standard mixtures the advantages of this ultrathin‐layer technique in terms of more convenient handling, cost and time are demonstra

ISSN:0173-0835    

DOI:10.1002/elps.1150050609

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

年代:1984

数据来源: WILEY

摘要:

AbstractThis paper describes the visualization of the low density lipoprotein (LDL) receptor by ligand blotting technique using an anti‐LDL‐enzyme linked immunosorbent assay (ELISA). Solubilized membrance proteins from bovine adrenal cortex are subjected to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, transferred to nitrocellulose paper and incubated with LDL. The combination of peroxidase‐con‐jugated second antibody and 4‐chloro‐naphthol/H2O2allows rapid development of colored bands where LDL was bound to the receptors. Saturation of the LDL binding to the LDL receptor can be demonstrated. This highly sensitive and rapid ELISA avoids the need for continuous supplies of iodinated

ISSN:0173-0835    

DOI:10.1002/elps.1150050610

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

年代:1984

数据来源: WILEY

摘要:

AbstractUltrathin‐layer isoelectric focusing of the urinary alanine aminopeptidase in 100 μm agarose gels resulted in a better separation of the isoenzymes and proteins than in polyacrylamide gels. Although miniature isoelectric focusing over a 4 cm separation distance proved excellent for screening purposes, more details were observed in gels with a 10 cm separation distance. Optimum enzyme visualization was achieved with a modified overlay technique employing a 0.2% agar with substrate instead of the conventional 1 % agar overlay. After incubation the diluted agar overlay may be easily washed off without damaging the running g

ISSN:0173-0835    

DOI:10.1002/elps.1150050611

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

年代:1984

数据来源: WILEY