title: Rapid preparation of peroxidase: anti-peroxidase complexes for immunocytochemical use.
abstract: Soluble immune complexes of horseradish peroxidase and antibody to peroxidase (PAP) have been widely used in the 'unlabelled antibody' method for the immunocytochemical detection of cellular antigens. This paper describes a simple and rapid method for preparation of these complexes by column chromatography of a mixture of the enzyme and the IgG fraction of antiperoxidase antiserum on Sephacryl S-200. The material eluting in the void column consists of stable soluble PAP complexes, with a molar peroxidase: antiperoxidase ratio of 0.8 and a molecular weight of approximately 400,000. When tested immunocytochemically this material gives identical results to those obtained with conventionally prepared PAP.
mesh_terms: D000818:Animals; D000936:Antigen-Antibody Complex; D002850:Chromatography, Gel; D005455:Fluorescent Antibody Technique; D006041:Goats; D006735:Horseradish Peroxidase / Q000276:immunology*; D006801:Humans; D005779:Immunodiffusion; D010544:Peroxidases / Q000276:immunology*; D011817:Rabbits; D013552:Swine
chemical_list: D000936:Antigen-Antibody Complex; D006735:Horseradish Peroxidase; D010544:Peroxidases