Preparation of peroxidase: antiperoxidase (PAP) complexes for immunohistological labeling of monoclonal antibodies.
The production of mouse peroxidase:antiperoxidase (PAP) complexes suitable for immunohistological use in conjunction with monoclonal antibodies is described. Three approaches were explored: 1) production of conventional polyclonal PAP complexes; 2) conversion of rabbit PAP to "pseudo-mouse PAP" by incubation with monoclonal mouse anti-rabbit immunoglobulin; 3) formation of PAP complexes from monoclonal mouse antiperoxidase. PAP complexes prepared by the latter technique gave the best immunohistological labeling reactions, being stable on storage and compatible with a wide range of human monoclonal antibodies. Gel filtration revealed that monoclonal PAP is of lower molecular weight than conventional PAP complexes (fulfilling theoretical predictions based on the monospecificity of monoclonal antibodies).
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006735:Horseradish Peroxidase / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D007146:Immunoglobulin lambda-Chains / Q000032:analysis; D051379:Mice; D008970:Molecular Weight; D010544:Peroxidases / Q000276:immunology*; D013194:Staining and Labeling
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007146:Immunoglobulin lambda-Chains; D006735:Horseradish Peroxidase; D010544:Peroxidases