A reliable method for simultaneous demonstration of two antigens using a novel combination of immunogold-silver staining with immunoenzymatic labeling

Please always quote using this URN: urn:nbn:de:bvb:20-opus-31092
  • We have developed a reliable and sensitive immunohistochemical staining technique which allows the simultaneous demonstration of two different antigens expressed in or on the same cell (referred to as mixed labeling), together with the evaluation of the general histopathological appearance of the tissue. The staining procedure combines a three-step (streptavidin-biotin) immunogold-silver staining (IGSS) with a three-step immunoenzymatic labeling. For this purpose, we investigated the compatibility ofIGSS with various substrates of peroxidase orWe have developed a reliable and sensitive immunohistochemical staining technique which allows the simultaneous demonstration of two different antigens expressed in or on the same cell (referred to as mixed labeling), together with the evaluation of the general histopathological appearance of the tissue. The staining procedure combines a three-step (streptavidin-biotin) immunogold-silver staining (IGSS) with a three-step immunoenzymatic labeling. For this purpose, we investigated the compatibility ofIGSS with various substrates of peroxidase or alkaline phosphatase (AP). Highly reliable and discernible mixed labeling was achieved only after iniriallabeling with IGSS followed by AP labeling using the substrates naphthol AS-MX phosphate/Fast Blue or naphthol AS-HI phosphate/New Fuchsin, respectively. To ensure utmost specificity, we applied FlTC-conjugated mouse monoclonal antibodies and rabbit anti-FlTC immunoglobulins visualized by AP-labeled immunoglobulins and the respective substrate in a final step. This novel approach provides an excellent means for demonstration of immunocompetent cells and unequivocal determination of the percentage of specific cell subsets in infiltrated tissue. The advantages of this method, as compared with double immunofluorescence or double immunoenzymatic labeling, were investigated and are discussed. (J Histochem Cytochem 38:307-313, 1990)show moreshow less

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Metadaten
Author: Reinhard Gillitzer, Rudolf Berger, Heidrun Moll
URN:urn:nbn:de:bvb:20-opus-31092
Document Type:Review
Faculties:Medizinische Fakultät / Institut für Molekulare Infektionsbiologie
Language:English
Year of Completion:1990
Source:In: J. of Histochemistry and Cytochemistry (1990) 38, 307-313.
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Tag:Immunohistochemistry; Immunogold-silver staining; FITC- anti-FITC system; Leucocyte subpopulations; Two-color staining
Release Date:2009/08/28