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. Sequencing grade modified trypsin is recombinant porcine trypsin modified by reductive methylation, rendering it resistant to proteolytic digestion.
Applications
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.
Enzyme Commission Number
EC 3.4.21.4
Activity
>4500 USP units/mg protein
Unit Definition
One USP unit of trypsin activity will produce a Delta A253 of 0.003 per minute in a reaction volume of 3.0ml at pH7.6 and 25°C, with BAEE as a substrate (1cm light path).
Storage
1. Sequencing Grade Modified Recombinant Trypsin lyophilized should be stored under 2°C-8°C in sealed container. It is stable within 24 months.After dissolved, it should be stored under -70°C, It is stable within 24 months.
2. Above 95% activity is remained after 5 times repeated freezing and thawing.
3. Above 90% activity is remained after kept under 4°C or 25°C for 24h.
4. A 0.05 mg/ml solution of sequencing grade modified recombinant trypsin in 50mM NH4HCO3 is retained above 95% after a 3 hours incubation at 37 °C . For long-term such as 20hours incubation, 1mM CaCl2 is recommended to be contained.
Preparation Instructions
1. Dilute recombinant modified trypsin with 50mM HAc if needed. When used, dilute it with 50mM NH4HCO3 or pH7.0-8.0 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 pH 7.0-8.0.
2. When using this product, thaw it at room temperature, and mix gently before use.
3. No activity lost when freeze-thaw 5 cycles.
Synonyms
α-trypsin; β-trypsin; cocoonase; parenzyme; parenzymol; tryptar; trypure; pseudotrypsin; tryptase; tripcellim; sperm receptor hydrolase; Alpha-trypsin; Beta-trypsin; EC 3.4.21.4; Trypsin; Acetyltrypsin; Trypsin, methylation modified; Methylation modified Trypsin