Detection of proteolytic enzymes in agar electrophoresis

An agar substrate containing hyaluronic acid and horse serum for the detection of hyaluronidase activity following electrophoresis was found suitable for the detection of proteolysis. The substrate was refined for the detection of only proteolysis by elimination of the hyaluronic acid and substitution of bovine serum albumin in place of the serum. The refined substrate revealed two bands of proteolytic activity in papain. The refined substrate also revealed weak proteolytic activity in a “chromatographically pure” preparation of hyaluronidase. The method should be adaptable for the detection of multiple electrophoretic forms of several proteolytic enzymes.