摘要:

AbstractA digital simulation of the behavior of amphoteric sample components in moving steady state boundaries is presented. Complete computer simulation data, including profiles of concentration, conductivity and pH as functions of time, are given for both cationic and anionic electrolyte configurations which incorporate one amphoteric sample constituent. The condensation of ampholytes in steady state moving boundaries is shown to proceed via an isotachophoretic mechanism and not by isoelectric focusing. Mobility (velocity) relationships necessary for sample components to form steady state zones are discussed.

ISSN:0173-0835    

DOI:10.1002/elps.1150070902

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

年代:1986

数据来源: WILEY

摘要:

AbstractBrief alkali treatment of nitocellulose‐bound protein markedly enhanced subsequent staining of protein with India ink or colloidal gold but not Amido Black. Following alkali treatment, India ink staining approaches colloidal gold staining in sensitivity and exceeds the latter method in clarity and economy. Furthermore, apparent protein‐to‐protein variation is reduced, allowing the method to be used for sensitive staining of protein transfers. Simple, high sensitivity staining of nitrocellulose‐bound protein using India ink can be achieved after alkali treatment in less than

ISSN:0173-0835    

DOI:10.1002/elps.1150070903

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

年代:1986

数据来源: WILEY

摘要:

AbstractHighly acidic proteins are difficult to study using conventional electrofocusing and electrophoretic techniques. For example, the lysosomal enzymes from the cellular slime moldDictyostelium discoideumhave isoelectric points (pI) around 3–4 due to extensive phosphorylation and sulfation. To assess differences in anionic modifications among these lysosomal enzymes, we required methods that resolved acidic proteins on the basis of their net charge. The separation procedures needed to be rapid and convenient, allowing parallel analysis of a large number of samples. Using these criteria, we developed two methods, isoelectric focusing in horizontal agarose gels and native electrophoresis in vertical polyacrylamide gels, which are more useful than other available techniques. Since the mechanism by which each system separates proteins is different, both methods give important information about acidic protein

ISSN:0173-0835    

DOI:10.1002/elps.1150070904

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

年代:1986

数据来源: WILEY

摘要:

AbstractPurified human blood clotting factor IX, although homogeneous by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, showed two peaks when analyzed by crossed immunoelectrophoresis in the presence of calcium. Differential affinity of lectin from wheat germ (WGA) for the two forms of factor IX, detected by crossed affino‐immunoelectrophoresis, demonstrated that carbohydrate moieties are involved in factor IX heterogeneities. The slow migrating peak has a higher affinity for WGA (Kd= 5.26 × 10−7M) than the fast migrating peak (Kd= 1.29 × 10−5M). This study emphasizes the usefulness of lectins as tools to assess homogeneity and integrity of glyc

ISSN:0173-0835    

DOI:10.1002/elps.1150070905

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

年代:1986

数据来源: WILEY

摘要:

AbstractWe have improved the Perrie and Perry method (Biochem. J.1970,119, 31–38) of urea‐electrophoresis by the addition of a stacking gel containing 8.5Murea and increased buffer capacity of the glycerol‐containing separation gel. The system has been successfully applied to the analysis of myosin phosphorylation levels not only in myosin and actomyosin, but also in whole muscle samples dissolved in urea. By subjecting the samples to a simple centrifugation and filtration step, all myosin light chains including the unphosphorylated and phosphorylated species as well as calmodulin and troponin could be identified. With a sensitivity of about 5 pmols the procedure described eliminates the need for the time‐consuming two‐dimensional electrophoresis which is commonly used to determine phosphorylation levels in muscl

ISSN:0173-0835    

DOI:10.1002/elps.1150070906

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

年代:1986

数据来源: WILEY

摘要:

AbstractMurine Cu, Zn‐superoxide dismutase (SOD) from erythrocytes has three isozymes, SOD‐1A (pI6.4), B (pI5.7) and C (pI5.0). It has been suggested that SOD‐1A is formed epigenetically by asymmetric modification of SOD‐1B where as SOD‐1C is produced by treatment of superoxide dismutase with hydrogen peroxide. On two‐dimensional gel electrophoretic analysis, each isozyme shows two forms, easily permuted into each other, and hence supposed to be in equilibrium. The zymogram patterns of murine liver, kidney, heart and brain are the same as those in erythrocytes although SOD activity levels are different in th

ISSN:0173-0835    

DOI:10.1002/elps.1150070907

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

年代:1986

数据来源: WILEY

摘要:

AbstractA simple and inexpensive system of a power supply for protein blotting is described. The blots are run using a 12 V car battery charger which is modified for laboratory use. Under these conditions even high molecular weight proteins of 300 000 can be completely transferred to the nitrocellulose paper in less than 5 h and efficiently immunostained.

ISSN:0173-0835    

DOI:10.1002/elps.1150070908

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

年代:1986

数据来源: WILEY

摘要:

AbstractSimple detection cells for on‐line UV absorbance and conductivity detection in capillary zone electrophoresis are described. The UV detection system employs a commercial photometer and optical fibers which are in direct contact with the outer walls of the separation capillary and conduct the light beam perpendicularly across the migrating zones. The conductivity detection cell consists of platinum wires protruding the wall of the separation capillary and thus in direct contact with the electrolytes inside. The electronic system is adopted from a commercially available conductivity detector for isotachophoresi

ISSN:0173-0835    

DOI:10.1002/elps.1150070909

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

年代:1986

数据来源: WILEY

摘要:

AbstractAn electrolyte system involving 2‐pyridinecarboxylate (Pic) buffer as the leading and formic acid as the terminating electrolyte was suggested for cationic capillary isotachophoresis of bases with low mobilities. Parameters of the system were studied for concentrations of the leading K+ion of 1–30 mmol/L and for the Hpic/Kpic ratios of 3:1, 1:1 and 1:3 in the buffer. The effective mobility of the terminating H+ion ranged between 3 and 30 X 10−9m2V−1s−1depending on the composition of the electrolyte system. The applicability of the system (at optimum concentration of K+5 mmol/L and the HPic/Kpic ratio 1:1) to the analysis of bases was characterized by a zone existence diagram and demonstrated by the separation of a model mixture containing tetramethylammonium, tetraethylammonium, tris(hydroxymethyl)aminomethane, ephedrine, codeine, nortriptyline, 1,10‐phenanthroline, aminophenazone and 4‐am

ISSN:0173-0835    

DOI:10.1002/elps.1150070910

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

年代:1986

数据来源: WILEY

摘要:

AbstractUrine analysis by two‐dimensional gel electrophoresis with isoelectric focusing in immobilized pH gradients in the first dimension allows the loading of high sample volumes and/or of large amounts of salts. When applied to urinary protein analysis, it makes it possible to apply up to 1 mL of dialyzed, unconcentrated sample or up to 350 μL of unprocessed uri

ISSN:0173-0835    

DOI:10.1002/elps.1150070911

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

年代:1986

数据来源: WILEY