Protocol using Trypsin-ultra™, Mass Spectrometry Grade (P8101)
Example Trypsin-ultraTM reaction (40:1 substrate:trypsin)
5 μl protein sample (about 2 ug)
4.5 μl Milli-QTM water
10 μl 2x Trypsin-ultra reaction buffer
0.5 μl 100 ng/μl Tyrpsin-ultra
Two useful references for trypsin behavior and usage:
Proc., J.L. et al (2010) “A quantitative study of the effects of chaotropic agents, surfactants, and solvents on the digestion efficiency of human plasma proteins by trypsin.” J. Proteome Res. 9, 5422-5437.
Vandermarliere, E. et al (2013) “Getting intimate with trypsin, the leading protease in proteomics.” Mass Spectrometry Reviews 32, 453-465.