Sequencing Grade Modified Trypsin, Frozen

Trypsin is a serine protease that specifically cleaves at the carboxylic side of lysine and arginine residues. The stringent specificity of trypsin is essential for protein identification. Native trypsin is subject to autolysis, generating pseudotrypsin, which exhibits a broadened specificity including a chymotrypsin-like activity. Such autolysis products, present in a trypsin preparation, would result in additional peptide fragments that could interfere with database analysis of the mass of fragments detected by mass spectrometry. Sequencing Grade Trypsin has been manufactured to provide maximum specificity. Lysine r...

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Trypsin is a serine protease that specifically cleaves at the carboxylic side of lysine and arginine residues. The stringent specificity of trypsin is essential for protein identification. Native trypsin is subject to autolysis, generating pseudotrypsin, which exhibits a broadened specificity including a chymotrypsin-like activity. Such autolysis products, present in a trypsin preparation, would result in additional peptide fragments that could interfere with database analysis of the mass of fragments detected by mass spectrometry. Sequencing Grade Trypsin has been manufactured to provide maximum specificity. Lysine residues in the porcine trypsin have been modified by reductive methylation, yielding a highly active and stable molecule that is extremely resistant to autolytic digestion.

The specificity of the purified trypsin is further improved by TPCK treatment, which inactivates chymotrypsin. The treated trypsin is then purified by affinity chromatography. It is resistant to mild denaturing conditions such as 0.1% SDS, 1M urea or 10% acetonitrile and retains 50% of its activity in 2M guanidine HCl. The activity of trypsin is decreased when acidic residues are present on either side of a susceptible bond. If proline is at the carboxylic side of lysine or arginine, the bond is almost completely resistant to cleavage.

Sequencing Grade Modified Trypsin, Frozen, is supplied in convenient 20µg aliquots as a frozen liquid in 50mM acetic acid.

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

Features - Benefits

• TPCK Treatment Followed by Affinity Purification: Elimination of chymotrypsin activity enables distinct and consistent data.
• Stability: Ensured up to five freeze-thaw cycles.
• Reliable and Customer-Proven: Referenced in thousands of papers.

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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