摘要:

AbstractUnder certain, defined conditions the temperature‐conductivity relationship of electrolytes leads to instabilities in the temperature distribution of electrophoresis gels if these are not efficiently thermostatted. These instabilities manifest themselves in practice in two ways: (i) as unequal temperatures of two identical gels connected in parallel and (ii) as an uneven temperature distribution across the face of individual gels. The latter is a principal cause of the undesirable “smile” effect that occasionally bedevils RNA and DNA sequencing gels. A quantitative description of these instabilities is put forward and suggestions for their avoidance are

ISSN:0173-0835    

DOI:10.1002/elps.1150080802

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

年代:1987

数据来源: WILEY

摘要:

AbstractGangliosides are known to exist as monomers in organic solvents and to aggregate in aqueous systems with the formation of high molecular weight micelles. It is shown here that the ganglioside micelles can be rendered visible by incorporation of a lipophilic dye and analyzed by conventional polyacrylamide gel electrophoresis. Mixed micelles of gangliosides and commonly used detergents as well as interaction products of ganglioside micelles with albumin were thus visualized by polyacrylamide gel electrophoresis for the first time.

ISSN:0173-0835    

DOI:10.1002/elps.1150080803

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

年代:1987

数据来源: WILEY

摘要:

AbstractWe have used six different monoclonal and polyclonal antibodies, four different antigen preparations and two different detection systems to compare Western blotting with Coomassie Blue prestained gels (Jackson and Thompson,Electrophoresis, 1984,5, 35–42) to blotting of unstained gels with Amido Black or Fast Green post‐transfer staining of the nitrocellulose. Contrary to a recent report (Harperet al., Anal. Biochem., 1986,157, 270–274), in which post‐staining with Coomassie Blue was determined to severely inhibit immunoreactivity, we find using prestained Coomassie Blue gels to be extremely useful in most cases, although, rarely, inhibition of the immune reaction may take place. Post‐staining with Amido Black or Fast Green may also prove useful, but is not recommended since similar inhibition occurs, and there are the added disadvantages of not being able to “pre‐view” the gel pattern and not being able to use stored gels. Further studies indicated that the rare loss of immunoreactivity seen with Coomassie Blue prestained gels is most likely due to the necessary fixation step, and not the stain itself (as has been suggested), since the protein‐dye complex is dissociate

ISSN:0173-0835    

DOI:10.1002/elps.1150080804

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

年代:1987

数据来源: WILEY

摘要:

AbstractA simple and rapid one‐dimensional analytical isoelectric focusing method followed by immunoblotting is described which detects genetic and biochemical variation in apolipoproteins A‐I, A‐II, A‐IV and C‐II without prior ultracentrifugation and delipidation of plasma samples. A polipoproteins separated on an isoelectric focusing gel are sequentially transferred to two nitrocellulose membranes and pairs of apolipoproteins are identified on each membrane by immunological reactions. The specificity of the primary antibody and the sensitivity gained by use of a second, alkaline phosphatase conjugated anti‐IgG allows the detection of free apolipoproteins in serum or plasma. A pH gradient 4–6.5 is suitable to identify the four apoproteins tested from a single gel and there is little loss of isoprotein resolution in sequential blots. Only 2 μL of whole plasma is required and results on 50 samples can be obtained from a single horizontal gel

ISSN:0173-0835    

DOI:10.1002/elps.1150080805

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

年代:1987

数据来源: WILEY

摘要:

AbstractThe effect of recombinant human tumor necrosis factor (rH‐TNF) on the composition and natural killer (NK)‐activity of lymphocytes in the spleen of normal and tumor‐bearing CBA mice was investigated. The characterization of cell composition was based on the analytical separation of splenic subpopulations differing in electrophoretic mobility. In untreated tunor mice, an increase of the electrophoretically intermediate population was found at the final stage of tumor growth. Treatment with TNF depressed this intermediate cell population in the spleen of tumorbearing mice, the electrophoretic distribution of splenocytes approaching the normal pattern. Also in normal mice TNF treatment affected the electrophoretic pattern. Already the first injection of TNF induced an increase of the intermediate cell population in the spleen. Tumor‐bearing mice showed a decreased NK‐activity at the final stage. In normal but not in tumor‐bearing mice repeated injections of TNF seemed to depress the NK‐activity. Cell electrophoresis represents a useful method for evaluating the effect of TNF on the cell populations in the s

ISSN:0173-0835    

DOI:10.1002/elps.1150080806

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

年代:1987

数据来源: WILEY

摘要:

AbstractA procedure for the isolation of protein markers of epidermal differentiationin vitrois described. Human epidermal keratinocytes were cultured and radiolabelledin vitro. Fractionation was performed according to buoyant density (which reflects the degree of terminal differentiation) using Percoll density gradient centrifugation. Subpopulations of keratinocytes were characterised using light and electron microscopy, and proteins fractionated using high resolution two‐dimensional gel electrophoresis. Radio‐labelled proteins were detected using autoradiography and fluorography. Integral membrane proteins were characterised using Triton X‐114 phase shift extraction. Data from thisin vitrostudy were compared to silver stained gels of samples from intact epidermis (in vivo). We report quantitative differences between 14 specific protein moieties expressed in subpopulations of keratinocytes and identify some of these proteins. The differential expression of these protein markers and their possible use in the interpretation of the keratinocyte maturation pathway in cultured cells from patients with skin diseases are disc

ISSN:0173-0835    

DOI:10.1002/elps.1150080807

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

年代:1987

数据来源: WILEY

摘要:

AbstractA simplified procedure for the preparation of ultrathin polyacrylamide slab gels is reported. The molding cells consist of an unsilanized glass plate and methacrylate plate or acryl‐coated glass plate. Owing to the repelling properties of acrylate the ultrathin gels spontaneously detach from the acrylate surface on unfastening the molds, firmly adhering to the unsilanized glass. Gels as thin as 0.20–0.25 mm could be polymerized without resorting to silanized glass plates or polyester films. The procedure is especially suitable for sticky gels, such as highly cross‐linked polyacrylamide gels and gels with immobilized pH grad

ISSN:0173-0835    

DOI:10.1002/elps.1150080808

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

年代:1987

数据来源: WILEY

摘要:

AbstractA micromethod of discontinuous polyacrylamide gel electrophoresis is described for phenotyping human haptoglobin in bloodstains. Pretreatment of samples is not required. This procedure is easy, fast and can detect haptoglobin phenotypes in 0.02 μL blood in experimentally produced stains

ISSN:0173-0835    

DOI:10.1002/elps.1150080809

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

年代:1987

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150080810

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

年代:1987

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150080811

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

年代:1987

数据来源: WILEY