Detection of elastase activity with a zymogram method after isoelectric focusing in polyacrylamide gel.
A zymogram method for detecting elastase activity following isoelectric focusing in polyacrylamide gel is described. After enzyme activity has been visualized, the gel itself is available for protein staining and for analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis in second dimension. The zymogram method is suitable for detecting microgram amounts of elastase and has one step only. It can be used with the purified enzyme as well as with crude extracts of tissue containing elastases showing activity toward succinyl-(Ala)3-p-nitroanilide. By this method a major component of elastase in both porcine and rat pancreas was detected. In addition, two forms of elastase with isoelectric points of 8.2 and 8.8, respectively, were identified in rat leukocyte extracts.
D000362:Agar; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D006098:Granulocytes / Q000201:enzymology; D007525:Isoelectric Focusing / Q000379:methods; D008832:Microchemistry; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology; D010196:Pancreatic Elastase / Q000032:analysis*; D051381:Rats; D013552:Swine
D000362:Agar; D010196:Pancreatic Elastase