Assay for the specificity of monoclonal antibodies in crossed immunoelectrophoresis.
A method is described based on crossed immunoelectrophoresis of a complex antigen mixture in agarose gel followed by incubation of the gel with the monoclonal antibody. The bound monoclonal antibody is detected by the use of a secondary enzyme-labelled antibody. Using this technique we have been able to identify the precipitate arc in crossed immunoelectrophoresis of major histocompatibility complex (MHC) class I molecules in a mixture of all detergent solubilized cell membrane molecules by means of a monoclonal antibody, the specificity of which was known independently to be against MHC class I molecules. In other experiments using the same technique we demonstrated the reaction of a monoclonal antibody specific for chicken Ig light chains.
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000961:Antilymphocyte Serum / Q000276:immunology; D001667:Binding, Competitive; D001798:Blood Proteins / Q000502:physiology; D011232:Chemical Precipitation; D002645:Chickens; D006649:Histocompatibility Antigens / Q000032:analysis / Q000276:immunology; D007123:Immunoelectrophoresis, Two-Dimensional; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits
D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum; D001798:Blood Proteins; D006649:Histocompatibility Antigens