DETECTION OF HEMOPROTEIN PEROXIDASE-ACTIVITY ON POLYVINYLIDENE DIFLUORIDE MEMBRANE

The feasibility of detecting hemoproteins after electroblotting was examined. Hemorproteins were subjected to lithium dodecyl sulfate-polyacrylamide gel electrophoresis (LDS-PAGE) and then electroblotted and peroxidase activity was detected with 3,3′,5,5′-tetramethylbenzidine. The sensitivity and specificity of tetramethylbenzidine staining of LDS-PAGE gels was retained when proteins were electroblotted. Subsequent staining of the membrane with Coomassie blue R-250 revealed a protein pattern similar to that in the polyacrylamide gel. Thus electroblotting of hemoproteins does not affect resolution of the electrophoretic pattern and heme-associated peroxidase activity. Additionally, the ability to stain hemoproteins on polyvinylidene difluoride membranes offers the advantage of utilizing the same membrane for further biochemical and immunological characterizations. © 1990.