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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 642 (1991), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A method has been developed to co-localize signals for antigen and DNA using a desktop microcomputer system (computer assisted signal co-localization). Antigens were detected by standard immunohistochemical methods and DNA was detected by non-isotopic in situ hybridization (NISH). Using this method, NISH signals can be precisely located in cells with well-preserved morphology captured by computer. The removal of the first immunohistochemical reaction products and reagents eliminates possible interference with hybridization and non-specific binding to the probe; therefore the sensitivity of the original NISH method remains. The captured NISH signals can be converted to any other colour which contrasts with the immunostaining. We have used detection of Epstein-Barr virus (EBV) and keratins as a model system. This method is straightforward, and with necessary modifications, will be applicable to any type of combined immunohistochemistry and in situ hybridization technique for simultaneous detection of antigen and nucleic acids or two types of nucleic acids in the same cells.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A procedure for the isolation of protein markers of epidermal differentiation in vitro is described. Human epidermal keratinocytes were cultured and radiolabelled in 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 this in vitro study 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 discussed.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A procedure is described for the detection of keratinocyte specific proteins. Fibroblasts and keratinocytes were isolated from human skin and radiolabelled in vitro. Samples were separated by two-dimensional polyacrylamide gel electrophoresis to compare the proteins synthesised by the different types of cultured skin cells. Dual label autoradiography of samples radiolabelled with [35S]methionine and [75Se]selenomethionine was used to identify keratinocyte specific proteins. We report 45 keratinocyte-specific components and identify some of these proteins. The differential expression of these proteins and their relevance to epidermal differentiation are discussed.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
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