Sequencing Grade Modified Trypsin, Used for Peptide Fingerprinting Analysis CAS 9002-07-7

Sequencing Grade Modified Trypsin, Used for Peptide Fingerprinting Analysis CAS 9002-07-7

Sequencing Grade Trypsin

Catalog Number:SRT02
EC:3.4.21.4

Source:Expressed in E. Coli, methylation modified.

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 sequencing grade modified trypsin is recombinant porcine trypsin modified by reductive methylation, rendering it resistant to proteolytic digestion.

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.

Reconstitution

Following reconstitution in 10 mM HCl, Trypsin, recombinant, proteomics grade is stable at -15 to -25 °C for up to one month. After first reconstitution the product must be stored in appropriate aliquots to avoid repeated freezing and thawing.

Specificity

Serine endopeptidase, hydrolyzing specifically proteins and peptides at the carboxy side of the basic amino acids Arg and Lys. Amide and ester bonds of Arg and Lys are also cleaved.

Application

Trypsin recombinant, Proteomics Grade, is used to specifically digest proteins separated by 2D gel electrophoresis or liquid chromatographic methods during sample preparation for mass spectrometry.