Trypsin Lys-C Sequencing Grade Trypsin for Mass Spectrometric Analysis

Trypsin Lys-C Sequencing Grade Trypsin for Mass Spectrometric Analysis

Sequencing Grade Trypsin

Description

Trypsin specifically hydrolyzes peptide bonds at the carboxyl side of lysine and arginine residues. Recombinant trypsin is free of any other proteases activities, and TPCK is unnecessary and not contained. Unmodified trypsin is subject to auto-proteolysis, generating fragments that can interfere with protein sequencing or HPLC/MS peptides analysis. YaxinBio’s sequence grade modified trypsin is recombinant porcine trypsin modified by reductive methylation, rendering it resistant to proteolytic digestion.

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.

Main Features

Unit Definition

One USP unit of trypsin activity will produce a Delta A253 of 0.003 per minute in a reaction volume of 3.2ml at pH7.6 and 25℃, with BAEE as a substrate (1cm light path).

Application

Protein digests for peptide mapping applications or protein identification by peptide mass fingerprinting or MS/MS spectral matching. It is suitable for digestion reactions in-solution or in-gel.

Recommend Usage

1.Dilute recombinant modified trypsin with 50mM HAc if needed. When used, dilute it with 50mM NH₄HCO₃ or pH7-8 buffers directly. 1mM CaCl₂ is recommended to be contained in digestion buffer. The ratio to aimed protein is 1:20 to 1:100 (w/w), the optimum pH is pH7-8.

2. When using this product, thaw it at room temperature, and mix gently before use.

3. No activity lost when freeze-thaw 5 cycles.

Storage Instructions

1. The solution should be stored under -70℃, It is stable within 24 months.

2. Above 95% activity remained after 5 times repeated freezing and thawing.

3. A 0.05 mg/ml solution of sequencing grade modified recombinant trypsin retained above 95% after a 3 hours incubation at 37 ℃ in 50mM NH₄HCO₃. For long-term such as 20hours incubation, 1mM CaCl₂ is recommended to be contained.