Hapten-mediated identification of cell membrane antigens using an anti-FITC monoclonal antibody.
A monoclonal anti-FITC antibody (F4/1) was produced and demonstrated to be specific for both the free and protein-conjugated (either soluble or cell-bound) form of fluorescein, or carboxyfluorescein. When mouse thymocytes were labelled with a novel fluorescein derivative 5(6)-carboxyfluorescein succinimidyl ester (CFl-NSE), the incorporation of fluorescein was predominantly membrane-bound as demonstrated immunohistochemically. The coupling of CFl-NSE to cells displays a random distribution pattern as shown by immunoblotting of cell extracts prepared by detergent solubilization of CFl-NSE-labelled thymocytes. In addition, the Thy-1.2 antigen immunoprecipitated from a CFl-NSE-labelled thymocyte lysate with a rat monoclonal antibody (Mab) could be detected using the anti-FITC Mab. The molecular weight of the immunoprecipitated material could be estimated immediately by reference to the FITC-labelled molecular weight markers electrophoresed simultaneously.
Balogh
P
P
Immunological and Biotechnological Laboratory, University Medical School of Pécs, Hungary.
Szekeres
G
G
Németh
P
P
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D019793:Fluorescein; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006241:Haptens; D006822:Hybrid Cells; D015151:Immunoblotting; D007150:Immunohistochemistry / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests; D051381:Rats; D013194:Staining and Labeling; D013950:Thymus Gland / Q000166:cytology*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D005452:Fluoresceins; D006241:Haptens; C024098:6-carboxyfluorescein; D016650:Fluorescein-5-isothiocyanate; D019793:Fluorescein