[Comparison of conditions for immunodetection of antigens using monoclonal antibodies after electrotransfer on nitrocellulose membrane].
Solubilized human sperm proteins are separated by SDS-polyacrylamide gel electrophoresis under reducing conditions, electrophoretically blotted onto a nitrocellulose sheet and their antigenicity probed with 5 monoclonal antibodies. Bound proteins are exposed to different nonionic detergents and a comparative study is reported. The use of 0.05% Tween 20 or 0.05% Nonidet P-40 in the blocking and washing buffers and in the immunoprobes causes variations in the patterns of 3 monoclonal antibodies. Washing buffers containing either 0.1% of detergent (Tween 20, Nonidet P-40, Triton X-100) or different concentrations (0.05, 0.1, 0.4%) of Tween 20 remove prebound proteins from the nitrocellulose membrane. Nitrocellulose strips processed with different detergents lack different immunoreactive bands. The 5 monoclonal antibodies recognize degraded antigens and endogeneous proteolysis changes the antibody patterns.
D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis*; D002021:Buffers; D002845:Chromatography; D003101:Collodion; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007120:Immunochemistry; D008297:Male; D051379:Mice; D013094:Spermatozoa / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D002021:Buffers; D003902:Detergents; D003101:Collodion
Comparaison des conditions d'immunodétection d'antigènes à l'aide d'anticorps monoclonaux après électrotransfert sur membrane de nitrate de cellulose.