Immunohistochemical application of monoclonal antibodies against myelin basic protein and neurofilament triple protein subunits: advantages over antisera and technical limitations.
Four monoclonal antibodies against guinea pig myelin basic protein (MBP), and four against subunits of bovine neurofilament triplet proteins (NF) were produced and their activity determined by enzyme-linked immunosorbant assay. The specificity and cross-reactivity of these eight monoclonal antibodies and one heterologous antiserum against each of the two central nervous system (CNS) antigens were examined in a histological study using the immunoperoxidase, antibody sandwich technique in rat and human brain tissue. Tissue sections were prepared from paraffin-embedded or fresh brain tissue that had been fixed with one of five different fixatives. The resulting immunoperoxidase labeling was then graded for intensity and examined for artifacts. One monoclonal antibody against MBP and one against NF resulted in labeling that was superior to that given by each of the antisera against their respective antigens. Of the five fixatives tested, a mercuric chloride-formalin solution gave the best preservation of these two antigens in rat and human brain tissue. The mercuric chloride-formalin solution was found to be superior to the other fixatives when immersion fixation was used, and was especially optimal when brains were perfused fixed. Three artifacts were encountered among the various antibody-fixative combinations that produced erroneous, but seemingly specific staining of Purkinje cells, neurons and axons, or astrocytes.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D001369:Axons / Q000032:analysis / Q000276:immunology; D001923:Brain Chemistry; D003712:Dendrites / Q000032:analysis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006651:Histocytochemistry; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D007381:Intermediate Filament Proteins / Q000276:immunology*; D008627:Mercuric Chloride; D008628:Mercury; D004676:Myelin Basic Protein / Q000276:immunology*; D016900:Neurofilament Proteins; D011689:Purkinje Cells / Q000032:analysis / Q000276:immunology; D051381:Rats; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D007381:Intermediate Filament Proteins; D004676:Myelin Basic Protein; D016900:Neurofilament Proteins; D008627:Mercuric Chloride; D008628:Mercury