Enhanced Protein Mass Spectrometry Analysis with Trypsin/Lys-C Mix Poster

Abstract

Sergei Saveliev, Ethan Strauss, Mike Rosenblatt and Marjeta Urh
Promega Corporation, Madison, WI

Efficient proteolysis is a major requirement for protein mass spectrometry analysis. Incomplete digestion has multiple ramifications including decreased number of identified proteins, compromised analytical reproducibility and protein quantitation, etc. Trypsin is one of the most robust proteases and is characterized by efficient proteolysis. However, even with trypsin, digestion is rarely complete. In a typical trypsin digest, 15-30% of sites have missed cleavages. In this study we show that, by supplementing trypsin with Lys-C, we eliminate majority of trypsin missed cleavages and substantially boost proteolytic efficiency.