Isolation of molecules recognized by monoclonal antibodies and antisera: the solid phase immunoisolation technique.
A simple technique for the isolation of antigens recognized by antisera and monoclonal antibodies has been developed. This method, the solid-phase immunoisolation technique, employs the protein-binding properties of polyvinylchloride microtiter plates. Antibodies are adsorbed to the plates either directly or via an anti-immunoglobulin reagent. Antigen is then placed in the wells, and allowed to adsorb to the antibody. The well is washed, and the antigen is then eluted with a denaturing electrophoresis sample buffer for one- or two-dimensional analysis. The solid-phase immunoisolation technique has been used to isolate a variety of cell membrane antigens with high signals and low backgrounds. The ease of the procedure and the high signal-to-noise ratio make this method preferable to the use of a staphylococcal adsorbent for many applications.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000302:isolation & purification*; D000941:Antigens / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera / Q000276:immunology; D007122:Immunoelectrophoresis; D007525:Isoelectric Focusing; D008722:Methods; D051379:Mice; D011143:Polyvinyl Chloride
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens; D007106:Immune Sera; D011143:Polyvinyl Chloride