A direct antigen-binding assay to screen hybridoma supernatants.
A simple and convenient method of directly assaying hybridoma supernatants for the desired monoclonal antibodies is described which obviates the need for labeled second or third antibody conjugates. Culture supernatants (1-5 microliters) were directly spotted onto a nitrocellulose sheet, and additional protein binding sites blocked with bovine serum albumin and incubated with enzyme-labeled, radioactive, or fluorescent antigen. Positive hybridoma supernatants were identified after washing and detection of bound antigen by appropriate means.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000941:Antigens / Q000276:immunology*; D001710:Biotin / Q000276:immunology; D003101:Collodion; D004798:Enzymes / Q000276:immunology; D006241:Haptens / Q000276:immunology; D006825:Hybridomas / Q000276:immunology*; D007118:Immunoassay / Q000379:methods*; D010544:Peroxidases / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D004798:Enzymes; D006241:Haptens; D001710:Biotin; D003101:Collodion; D010544:Peroxidases