Analysis of chemical and biochemical properties of membrane molecules in situ by analytical flow cytometry with monoclonal antibodies.
Methods are described for analysing chemical and biochemical properties of membrane antigens against which monoclonal antibodies are available. The methods are based on gentle manipulation of whole viable cells and quantitative flow cytometric fluorescence analysis of the effect of such treatment on monoclonal antibody binding. Proteolytic enzymes, glycosidases, inhibitors of biosynthesis and various mild chemical treatments have been used to derive information on the chemical nature of individual membrane antigens, their insertion in the membrane, and their turnover. The nature of the particular epitope detected by a monoclonal antibody may be probed in a similar way.
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000954:Antigens, Surface / Q000032:analysis*; D001402:B-Lymphocytes / Q000276:immunology; D002241:Carbohydrates; D002460:Cell Line; D005434:Flow Cytometry / Q000379:methods; D005455:Fluorescent Antibody Technique; D006026:Glycoside Hydrolases; D006801:Humans; D037102:Lectins; D013601:T-Lymphocytes / Q000276:immunology; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000954:Antigens, Surface; D002241:Carbohydrates; D037102:Lectins; D014415:Tunicamycin; D006026:Glycoside Hydrolases