title: Quantitative immunohistochemistry in the rat facial nucleus with [125I]-iodinated secondary antibodies and in situ autoradiography: non-linear binding characteristics of primary monoclonal and polyclonal antibodies.
abstract: Indirect immunohistochemistry is an important routine method in histology and histopathology. Here we have investigated the quantitative aspects of antibody binding to tissue sections, using a range of monoclonal and polyclonal antibodies and the regenerating rat facial nucleus as the experimental model. The in situ binding of primary antibodies was quantified using appropriate [125I]-iodinated secondary antibodies and quantitative autoradiography. The majority of primary antibodies revealed an apparently bell-shaped curve of in situ antibody binding, with the binding increasing up to a specific antibody concentration and then decreasing; similar data were also obtained with enzymatic immunohistochemistry. There was also a close correlation between the quantitative changes in antibody binding during the time course of facial nerve regeneration and those observed with enzyme histochemistry.
authors.affiliation: Department of Neuromorphology, Max-Planck-Institute for Psychiatry, Martinsried, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Raivich
authors.forename: J
authors.initials: J
authors.lastname: Gehrmańn
authors.forename: M B
authors.initials: MB
authors.lastname: Graeber
authors.forename: G W
authors.initials: GW
authors.lastname: Kreutzberg
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001345:Autoradiography; D001933:Brain Stem / Q000737:chemistry* / Q000276:immunology; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D008433:Mathematics; D051381:Rats; D017208:Rats, Wistar; D011951:Receptors, Complement / Q000276:immunology; D014746:Vimentin / Q000276:immunology
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D005904:Glial Fibrillary Acidic Protein; D015395:Histocompatibility Antigens Class I; D011951:Receptors, Complement; D014746:Vimentin