Application of avidin-biotin-peroxidase complex method with a monoclonal antibody to human Ia-like antigen in immunoelectron microscopy.
Monoclonal antibody (MAb) to human Ia-like (HLA-DR) antigen was applied with the avidin-biotin-peroxidase complex (ABC) immunostaining method to localize the Ia-like antigen at the electron microscopic level. Our results indicated that in human tonsils and adenoids fixed with 4-6% phosphate-buffered paraformaldehyde for 4-6 hr, sharply delineated electron-dense products of the antigen and antibody complex were detectable on the outer cell membranes of lymphoblasts, lymphocytes, reticular cells, and macrophages. In our study, the vibratome sections of the paraformaldehyde-fixed, pre-embedding immunostained tissues consistently showed more satisfactory morphology than frozen sections. The combined use of the anti-human Ia monoclonal antibody and the ABC procedure with paraformaldehyde fixation provides a simple and sensitive method to study at the ultrastructural level the Ia-like antigen-bearing cells, which are vital in the immune response.
D000234:Adenoids / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001360:Avidin; D001710:Biotin; D006681:HLA-D Antigens / Q000032:analysis*; D006684:HLA-DR Antigens / Q000032:analysis*; D006801:Humans; D007124:Immunoenzyme Techniques; D008854:Microscopy, Electron; D014066:Palatine Tonsil / Q000276:immunology
D000911:Antibodies, Monoclonal; D006681:HLA-D Antigens; D006684:HLA-DR Antigens; D001360:Avidin; D001710:Biotin