Production and study of antibody produced aganist rat cadmium thionein

Abstract

Cadmium binding protein (CdBP)^* has been isolated from rat liver and separated by DEAE ionexchange chromatography into its two forms: CdBP-1 and CdBP-2. An analysis of the purity of these two proteins has been made by disc gel clectrophoresis and slab gel electrophoresis. It was found that the first cadmium binding protein peak to elute from the DEAE separation, referred to as CdBP-1, contained significant amounts of impurities, whereas the second peak, referred to as CdBP-2, was electrophoretically pure. Rabbits were immunized with either monomeric CdBP or glutaraldehyde-trcated CdBP. Antisera from these rabbits have been tested for their ability to bind CdBP and ZnMt. Antigen binding capacity (ABC), double diffusion in gels and competitive binding assays were used to characterize the strength and specificity of the antisera. The anti-CdBP produced cross reacts completely with CdBP-1 and CdBP-2 as well as with ZnMt-A and ZnMt-B.