The determination of dissociation constants by affinity electrophoresis on Cibacron Blue F3G A-agarose-polyacrylamide gels

The application of copolymerized agarose-polyacrylamide gels as the support for immobilized Cibacron Blue F3G A is demonstrated for the analytical electrophoresis of proteins possessing an affinity for this dye. Bovine serum albumin was used as a model protein to develop this technique. The optimal conditions for preparing matrices are described. These conditions produce gels with suitable mechanical strength and which allow rapid electrophoresis of proteins. The dye-agarose-polyacrylamide gels permit the determination of dissociation constants. The ease of preparation of these matrices recommends them for a variety of quantitative analytical investigations.