A comparative study of avidin-biotin-peroxidase complexes for the immunohistochemical detection of antigens in neural tissue.
It has been suggested that the use of avidin-biotin immunohistochemical techniques for antigen detection in neural tissue produces nonspecific background staining. For this reason neural tissue was used to test the quality, sensitivity and specificity of four commercially available antibody detection kits which use avidin or streptavidin binding to biotin. Free-floating, thick-section immunohistochemistry on perfusion fixed rat central nervous system revealed variability among staining kits for all parameters analyzed under the same experimental conditions. The reagents from the Vector 'Elite' kit were the most sensitive and specific, and received the highest overall rating for quality. Most commercial products tested could be used at greater dilutions than those recommended by the manufacturers without compromising specific staining. No staining was evident when the primary and secondary antibodies were omitted. This suggests that nonspecific binding is unlikely to be due to endogenous ligands, charge or hydrophilic reactions between these tertiary complexes and the tissue sections.
Ellis
J
J
Department of Pathology, University of Sydney, Australia.
Halliday
G
G
D000818:Animals; D000906:Antibodies / Q000032:analysis; D000941:Antigens / Q000032:analysis*; D001360:Avidin; D001426:Bacterial Proteins; D001710:Biotin; D001921:Brain / Q000276:immunology* / Q000378:metabolism; D001923:Brain Chemistry; D007150:Immunohistochemistry / Q000191:economics / Q000379:methods; D008297:Male; D010320:Parvalbumins / Q000378:metabolism / Q000493:pharmacokinetics; D010544:Peroxidases; D051381:Rats; D017208:Rats, Wistar; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D019809:Streptavidin
D000906:Antibodies; D000941:Antigens; D001426:Bacterial Proteins; D010320:Parvalbumins; D011933:Reagent Kits, Diagnostic; D001360:Avidin; D001710:Biotin; D019809:Streptavidin; D010544:Peroxidases