Detection of proteolytic activity after isoelectric focusing in polyacrylamide gel

Abstract

δ-Chymotrypsin, subtilopeptidase and crude extracellular proteases from Pseudomonas aeruginosa have been separated by isoelectric focusing in thin layers of polyacrylamide gel. Ampholine was removed by soaking the gel in buffer, prior to overlaying with casein-containing agar to detect proteolysis. The technique described is simple and has a high resolving power. It should have general application for the analysis of enzymes.