Immunohistochemical detection of antibody in tissue sections of non-perfused and ex vivo-perfused organs using a tetrazolium alkaline phosphatase substrate.
We have used nitroblue tetrazolium (NBT) as a color reagent to localize antibody-bound alkaline phosphatase in frozen tissue sections. In the method described, NBT is reduced to a stable black diformazan reaction product that contrasts well with nuclear counterstains such as hematoxylin and stands out strongly in black and white photographs. We have found NBT to be a suitable color reagent for the alkaline phosphatase: anti-alkaline immunohistochemical technique. The reaction product also contrasts well with fast red and can therefore be used as second reagent for two color immunoenzyme studies. In this report, we describe a novel two color immunoenzyme method to assess the ex vivo binding of antibodies against Class II histocompatibility antigens in whole organs connected to a perfusion circuit.
D000469:Alkaline Phosphatase / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry / Q000379:methods*; D066298:In Vitro Techniques; D009580:Nitroblue Tetrazolium; D010179:Pancreas / Q000166:cytology / Q000201:enzymology*; D010477:Perfusion / Q000379:methods*; D051381:Rats; D011917:Rats, Inbred Lew; D013778:Tetrazolium Salts
D000911:Antibodies, Monoclonal; D013778:Tetrazolium Salts; D009580:Nitroblue Tetrazolium; D000469:Alkaline Phosphatase