Immunochemical detection of plasma proteins after two-dimensional electrophoresis

Abstract

A technique of immunochemical detection of plasma proteins on the slab gel after two-dimensional electrophoresis was described. The slab gel (4mm in thickness) was cut into 2 slices (2mm in thickness). One of the slices was stained with coomassie blue and the other was overlaid with antibody-containing agar layer. Bovine serum amine oxidase, as small amount as 0.5μg, could be detected by immunodiffusion followed by coomassie blue staining of the agar layer. Human myeloma IgA and human IgG were also detected by this technique. These results demonstrated the advantage of the two-dimensional technique which did not employ denaturing agent such as urea or SDS.