Characterization of Natural Inhibitors of Trypsin and Chymotrypsin by Electrophoresis in Acrylamide-Agarose Gels

Abstract

WE have detected natural inhibitors of enzymes m microgram quantities. In our method, after gel electrophoresis of a sample presumed to contain inhibitory activity, the slab of gel was incubated in a solution of the appropriate enzyme which then entered the gel by diffusion and formed a thin and homogeneous layer on its surface. After several minutes the gel was removed from the solution, allowed to stand until completion of the enzyme-inhibitor complex, and then transferred into a solution containing a chromogenic substrate for the enzyme used in the assay. In these conditions, the catalytic activity of the enzyme could be visualized, for the whole surface of the gel was stained, except for areas where the inhibitor was present.