A novel antibody overlay technique for two-dimensional immunoelectrophoresis.
A new antibody overlay technique for 2-dimensional immunoelectrophoresis (2-D-IEP) is described. After the first dimension electrophoresis of the antigen, the desired amount of antiserum is applied to the initial agarose layer and then evenly distributed over a defined surface of the gel with a PVC film. This modification of the conventional 2-D-IEP procedure makes it possible to perform tandem 2-D-IEPs comparing 2 antisera on the same gel plate, rocket IEPs where several antisera are compared by electrophoresis on the same gel plate, and 2-D-IEPs with an intermediate antiserum, avoiding the need to pour an intermediate gel. With this technique, 77 antigens have been demonstrated in a Candida albicans serotype A somatic antigen preparation. This is the first description of such a large number of immunoprecipitates on the same immunoplate.
D000818:Animals; D000908:Antibodies, Fungal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000946:Antigens, Fungal / Q000276:immunology; D002176:Candida albicans / Q000276:immunology; D011232:Chemical Precipitation; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007122:Immunoelectrophoresis / Q000379:methods*; D007123:Immunoelectrophoresis, Two-Dimensional / Q000379:methods*; D011817:Rabbits
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D007106:Immune Sera