Journal
PROTEOMICS
Volume 5, Issue 9, Pages 2319-2321Publisher
WILEY
DOI: 10.1002/pmic.200401268
Keywords
enzyme characterization; protein digestion; sample preparation; trypsin
Funding
- NIMH NIH HHS [R01MH59926] Funding Source: Medline
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Prior to analysis by mass spectrometry, protein samples are often digested. Maximizing the peptide yield from digestion can increase the number of peptides detected and the confidence in protein identification. To determine the optimal conditions for digestion, the Michaelis-Menten kinetic parameters for Promega sequencing grade modified trypsin were measured over a range of temperatures and pHs. The results indicate that an increase in digestion temperature above 37 degrees C, the temperature traditionally used in digestion methods, could offer an increase in peptides detected.
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