Detection of isoelectric focused antibody by autoradiography and hemolysis of antigen-coated erythrocytes. A comparison of methods.

Mouse anti-group A streptococcal carbohydrate (GAC) antibodies were detected in analytical isoelectric focusing (IEF) acrylamide gels by the hemolysis of GAC-coated sheep erythrocytes immobilized in agarose. This procedure is rapid and reproducible but, because it requires diffusion of antibody from the acrylamide gel into the agarose layer, results in poorer resolution than other methods of detection where antibodies can be fixed in the acrylamide gel after focusing, such as protein staining of purified antibodies or autoradiography of 125I-GAC bound to focused antibodies.