Detection of endoproteinases in polyacrylamide gels

✍ Scribed by Gary R. Petersen; Dr. James L. Van Etten

• Publisher: John Wiley and Sons
• Year: 1983
• Tongue: English
• Weight: 314 KB
• Volume: 4
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.1150040613

✦ Synopsis

Abstract

A simple and versatile method was developed for detecting endoproteinases by nondenaturing polyacrylamide gel electrophoresis. A vertical strip of the resolving gel is subjected horizontally to a second electrophoresis in the presence of a substrate protein and sodium dodecyl sulfate (SDS). The location of proteolytic activity in the original gel strip is revealed by the disappearance of substrate and the appearance of proteolytic products (due to limited proteolysis) in the second gel. The pattern of peptide fragments produced permits the identification of the protease responsible.