The IHC Select® Immunoperoxidase Secondary Detection System is intended for use with the CHEMICON IHC Select® Prediluted Primary Antibody reagents, which contain rabbit and mouse IgG for the qualitative identification of antigens by light microscopy in paraffin-embedded tissues. The IHC Select® Immunoperoxidase Secondary Detection System may also be used with other rabbit or mouse primary antibodies.
The IHC Select® Immunoperoxidase Secondary Detection System uses biotin-avidin-HRP complexed antibodies to detect rabbit or mouse IgG in the Primary Antibody. The primary antibody binds to specific antigens present in the specimen. Any excess antibody is removed by washing. Next, the Secondary Antibodies Reagent (conjugated to biotin) is added and will react with the primary antibody present on the tissue. Unbound antibody is removed by washing. The sample is then by incubated with the Streptavidin-HRP solution, which binds to the biotin-labeled secondary antibody present on the tissue. Unbound enzyme is removed by washing. The chromogenic development reagent, 3,3' diaminobenzidine (DAB Substrate), is then added and reacts with the HRP attached to the HRP-streptavidin-biotin-antibody complex. The HRP activity on the chromogenic substrate results in the deposit of brown to black insoluble precipitate at those antigenic sites containing the specific epitopes recognized by the primary antibody. Tissues processed in a variety of fixatives including neutral buffered formalin, B-5, and Bouin's may be used.