Biotinylation: an alternative to radioiodination for the identification of cell surface antigens in immunoprecipitates.
A method has been developed in which the conventional radioiodine label is replaced by non-radioactive biotin in studies involving the immunoprecipitation and analysis of cell surface antigens. The labelling reagent, d-biotinyl-N-hydroxysulfosuccinimide ester (NHSS-biotin), reacts preferentially with lysine residues in polypeptides and possibly also with free amino-groups on carbohydrates and lipids. The reagent can be used as a cell surface label, does not interfere with antigen-antibody interactions and allows labelled molecules to be detected with high sensitivity using streptavidin-peroxidase conjugates. The target antigens of a range of monoclonal antibodies to human cell surface components have been identified using this procedure.
Cole
S R
SR
Department of Microbiology and Immunology, University of Adelaide, Australia.
Ashman
L K
LK
Ey
P L
PL
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000032:analysis*; D000954:Antigens, Surface / Q000032:analysis*; D001710:Biotin / Q000031:analogs & derivatives*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007124:Immunoenzyme Techniques; D007938:Leukemia / Q000276:immunology*; D011863:Radioimmunoassay; D013388:Succinimides
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D013388:Succinimides; C053827:biotinyl-N-hydroxysulfosuccinimide ester; D001710:Biotin