ISSN:0173-0835    

DOI:10.1002/elps.1150050102

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

年代:1984

数据来源: WILEY

摘要:

AbstractRheophoresis,i.e.thin‐layer electrophoresis in particulate gels with controlled evaporation, combines in one experiment the chromatographic and the electrophoretic characteristics of macromlecules in such a way that they can be determined independently of each other. In this paper theory and experimental corroboration for molecular size determination are presented. We describe the migration velocity of any molecule as the sum of an electrophoretic, an electroosmotic and a rheophoretic component. The latter is the velocity originating from the liquid flow produced by evaporation. It is a function of the position along the gel axis and the cross‐section available to the molecule, but totally independent of the molecular charge. As a result of the position dependence, the rheophoretic component can be isolated from the two others and the available cross‐section determined by means of a simple plot. Thus, the chromatographic partition coefficient is readily calculated and the effective hydrodynamic radius of the molecule obt

ISSN:0173-0835    

DOI:10.1002/elps.1150050103

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

年代:1984

数据来源: WILEY

摘要:

AbstractGroup specific component (Gc) is known to interact with both vitamin D3metabolites and actin. The present study was undertaken to determine if analytical isoelectric focusing could form the basis of a simple and reliable method for discriminating native Gc and complexes with 25‐hydroxycholecalciferol (25(OH)D3) from those formed with actin and with both ligands. Increasing amounts of G‐actin and/or 25(OH)D3were added to purified Gc of both Gc1 and Gc2 phenotypes. Actin and 25(OH)D3interacted independently and simultaneously with native Gc, giving rise to three different complexes of increasing acidity: Gc‐25(OH)D3, Gc‐actin, and actin‐Gc‐25(OH)D3, which were clearly resolved from the native Gc protein and from each other. In addition, the inherent differences in microheterogeneity and polymorphism between the two major phenotypes examined (Gc1 and Gc2) were also retained in their respective complexes. Similar results were obtained upon addition of 25(OH)D3or actin to whole human serum, the bands corresponding to native or complexed Gc being recognized in this case by print immunofixation. These results indicate that complexes formed between Gc protein and both 25(OH)D3and actin can be clearly detected and resolved by electrophoretic

ISSN:0173-0835    

DOI:10.1002/elps.1150050104

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

年代:1984

数据来源: WILEY

摘要:

AbstractA widely applicable procedure for electrophoretic separation and quantification of proteins is described. Different colored markers of myoglobin were prepared by carbamylation and those with suitable isoelectric points (pI's) were isolated. Such markers were added to samples of serum and plasma which were examined by improved isoelectric focusing in agarose gel. This facilitated isolation of a certain transferrin with pI5.7. This protein was quantified in gel pieces by zone immunoelectrophoresis assay. The overall S.D. was<10%. Immunofixations were performed to check the quality of focusing and the accuracy of gel punching.

ISSN:0173-0835    

DOI:10.1002/elps.1150050105

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

年代:1984

数据来源: WILEY

摘要:

AbstractA simple technique has been developed for the electrophoretic extraction of milligram to microgram quantities of protein, under constant monitoring by dye line, in a concentrated form from polyacrylamide gels run under alkaline, neutral and acidic conditions. Typically, the extraction time from 5 gel slices per tube containing about 1.2 mg of bovine serum albumin or α‐chymotrypsinogen A was 3 h, and the maximum concentration of the recovered protein was 5 mg/ml with a total recovery of more than 90% in all cases. Higher protein concentrations (11 mg/ml) could be obtained by increasing the extraction time to 3.5 h or by using resins like glass beads. The alkaline extraction system has been applied to the purification of glutamyl‐tRNA synthetase up to homogeneity, and also to ferritin. This technique can also be used under reducing conditions and the scale of operation can be adjusted by the number of tubes or number of sl

ISSN:0173-0835    

DOI:10.1002/elps.1150050106

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

年代:1984

数据来源: WILEY

摘要:

AbstractProteins visualised by Coomassie Blue staining of two‐dimensional polyacrylamide gels of complex mixtures of brain proteins were blotted onto nitrocellulose sheets. The Coomassie Blue stain was transferred simultaneously and bound strongly to the nitrocellulose, giving a copy of the original gel pattern. Using specific antisera and a second antibody coupled to horseradish peroxidase, proteins in the original mixture could be detected on the nitrocellulose in quantities less than 25 ng. The Coomassie Blue and horseradish peroxidase stains were distinguishable by a suitable filter, allowing precise correlation of spots stained by Coomassie Blue with spots stained with the brown immunoperoxidase reaction product. The technique was used to identify related proteins in electrophoretograms of brain extracts from different animal

ISSN:0173-0835    

DOI:10.1002/elps.1150050107

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

年代:1984

数据来源: WILEY

摘要:

AbstractA simple apparatus and a convenient program for evaluation of two‐dimensional electrophoretograms have been developed. The hardware system comprises a charge‐coupled device (CCD) camera, four pages of frame memory (FM), a page of display memory (DM), a video‐monitor cathode‐ray tube (CRT), a digitizer (DIG) and two ports of parallel interfaces (PIO). A small desk‐top microcomputer is connected to the apparatusviathe interfaces. The system program named frame‐imagememory controller (FIMC) has been developed under an assembler of the microcomputer. FIMC is a package of subroutines, and utility programs for two‐dimensional densitometry can be written in BASIC language simply with the help of FIMC. Unevenness of basal gray level, caused by the nonuniformity of illumination in the camera's field of view, is eliminated from the pixel data by a method of blank subtraction. Background gray level, caused by protein staining procedures, is corrected by a method of local background subtraction. The two‐dimensional densitometry system, equipped with a CCD camera, is applicable not only to transparent gels but also to opaque membranes, and it shows a high reliability in optical measurement in the r

ISSN:0173-0835    

DOI:10.1002/elps.1150050108

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

年代:1984

数据来源: WILEY

摘要:

AbstractHigh resolution two‐dimensional electrophoresis combined with a sensitive silver stain has been used to study the proteins of normal human aqueous humour. The protein patterns of small, unconcentrated, individual samples and of pooled and concentrated aqueous humour samples were compared with plasma or serum from the same individual or with a normal serum. The protein maps of aqueous humour and serum showed great similarity. More than 30 groups of spots were visualized in aqueous humour after silver staining and 18 of them have been tentatively identified. Three polypeptide groups were observed which were not found in plasma or serum. The different clusters, which appeared in the region of high (>90 000), medium (50 000–45 000) and low (25 000–15 000) molecular weight polypeptides, remain unidentified. The most abundant cluster represents a molecular weight of approximately 4

ISSN:0173-0835    

DOI:10.1002/elps.1150050109

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

年代:1984

数据来源: WILEY

摘要:

AbstractNonspecific antibody binding either to nitrocellulose membrane sites or to transferred and immobilized proteins is eliminated (or minimized to insignificant levels) by treatment with a liquid gelatin preparation. This increases the specificity and sensitivity of nitrocellulose binding immunoassays, and allows incubation at 4°C which minimizes bacterial contamination and/or damage to biologically‐labile protei

ISSN:0173-0835    

DOI:10.1002/elps.1150050110

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

年代:1984

数据来源: WILEY

摘要:

AbstractIn a study of intersterility groups in the genusArmillaria, a scanning stage microscope photometer was used to obtain digitized images of photographs of isoelectric focusing patterns. Absorbance values were then mapped from the metric scale to the pH scale. Absorbance values in specified pH intervals were used as features in multivariate statistical procedures to characterize populations and to assign isolates to populations. Digital image analysis techniques were used to obtain an enhanced image of faint or poorly resolved bands.

ISSN:0173-0835    

DOI:10.1002/elps.1150050111

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

年代:1984

数据来源: WILEY