Sequencing Grade Modified Trypsin

Trypsin specifically hydrolyzes peptide bonds at the carboxylic sides of lysine and arginine residues. Unmodified trypsin is subject to autolysis, generating fragments that can interfere with protein sequencing, HPLC or mass spectrometry analysis of the peptides. In addition, autolysis can result in the generation of pseudotrypsin, which has been shown to exhibit an additional chymotrypsin-like specificity. Promega Trypsin has been modified by reductive methylation, rendering it extremely resistant to autolytic digestion. In functional stability tests, modified trypsin retains at least two times as much activity a...

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Trypsin specifically hydrolyzes peptide bonds at the carboxylic sides of lysine and arginine residues. Unmodified trypsin is subject to autolysis, generating fragments that can interfere with protein sequencing, HPLC or mass spectrometry analysis of the peptides. In addition, autolysis can result in the generation of pseudotrypsin, which has been shown to exhibit an additional chymotrypsin-like specificity. Promega Trypsin has been modified by reductive methylation, rendering it extremely resistant to autolytic digestion. In functional stability tests, modified trypsin retains at least two times as much activity as unmodified trypsin after a 3-hour incubation at 37°C.

The sequencing grade of modified trypsin has been further improved by TPCK treatment followed by affinity purification yielding a highly active and stable molecule. Sequencing Grade Modified Trypsin is provided as a lyophilized powder in convenient 20μg aliquots with a stability-optimized resuspension buffer. A protease:protein ratio of 1:100 to 1:20 (w/w) is recommended.

Recommended Reaction Buffer: 50mM NH₄HCO₃ (pH 7.8).

Features - Benefits

• Pure: TPCK treatment followed by affinity purification.
• Easy to Use: Resuspension buffer provided.
• Convenient: Five tubes of lyophilized product are provided.
• Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/

References

1. Keil-Dlouha, V.V. et al. (1971) FEBS Letters 16, 291–5.
2. Rice, R.H. et al. (1977) Biochem. Biophys. Acta 492, 316–21.

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