Recombinant trypsin is expressed and produced by recombinant Escherichia coli. Its amino acid sequence is identical to that of cationic trypsin derived from porcine pancreas. It is free of contaminating enzymes such as chymotrypsin; TPCK is neither contained nor required to inhibit interfering enzymatic activities. It features high activity and stringent digestion specificity.
Conventional trypsin is prone to autolysis, generating autolytic fragments that interfere with mass spectrometry results. Methylation modification and site-directed mutations eliminate its autolytic capacity, and consequently abolish pseudo-chymotrypsin activity arising from autodegradation.
Recommended Protocol
Dissolve or dilute the product with 50 mM HAc or 1 mM HCl. For digestion, further dilute it in 50 mM NH₄HCO₃ or buffer solutions at pH 7.0–8.0.
Add 1 mM CaCl₂ to the digestion buffer as recommended. The mass ratio of recombinant trypsin to target protein ranges from 1:20 to 1:100, with an optimal pH of 7.0–8.0.
Product Advantages
1. Sequence-grade pure protease: Excellent substrate specificity and superior stability.
2. Animal-origin-free: Manufactured via recombinant expression without any animal-derived raw materials, eliminating risks of exogenous viral contamination.
3. Consistent quality: Mass production enables stable continuous batch manufacturing with negligible inter-batch variability.
4. High purity & specific activity: Residual host cell protein levels are below the limits specified for biological products.
5. Lyophilized powder format: Convenient for storage and transportation.
Applications
Recombinant trypsin shares identical enzymatic properties with animal-derived trypsin and serves as a superior alternative thanks to the absence of contaminating enzymatic activities, enhanced stability, resistance to autolysis, and high catalytic activity. No non-specific cleavage fragments or autolytic byproducts are generated during digestion.
It is widely applied in site-specific protein digestion, protein sequencing, peptide mapping, proteomics research, and in-gel tryptic digestion of proteins separated by SDS-PAGE.
G2631632