title: Quantitative study of enzyme immunocytochemical reactions performed with enzyme conjugates immobilized on nitrocellulose.
abstract: The nitrocellulose binding assay was used for quantitative studies on the cytochemical reactions for the three enzymes most frequently used in immunocytochemistry. The results show a linear relationship between the amount of enzyme immobilized on nitrocellulose and the amount of the enzyme reaction product. The similar course of the formation of the reaction product after DAB/H2O2 staining for peroxidase immobilized on nitrocellulose and for immunoperoxidase labeled cells indicates a linear relationship between the amount of enzyme-coupled antibodies bound to cells and the amount of enzyme reaction product. Furthermore, a mild acid treatment for the abolition of endogeneous peroxidase activity in tissues and cells applicable to immunoperoxidase staining procedures is proposed.
mesh_terms: D015100:3,3'-Diaminobenzidine / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D003101:Collodion; D005696:Galactosidases / Q000032:analysis*; D006651:Histocytochemistry / Q000379:methods; D006735:Horseradish Peroxidase / Q000032:analysis*; D007700:Kinetics; D008264:Macrophages / Q000032:analysis; D051379:Mice; D010544:Peroxidases / Q000032:analysis*; D001616:beta-Galactosidase / Q000032:analysis*
chemical_list: D015100:3,3'-Diaminobenzidine; D003101:Collodion; D006735:Horseradish Peroxidase; D010544:Peroxidases; D000469:Alkaline Phosphatase; D005696:Galactosidases; D001616:beta-Galactosidase