Description

Trypsin (EC 3.4.21.4) is a serine protease that hydrolyzes peptide bonds at the C-terminal of lysine and arginine residues. Recombinant porcine trypsin is expressed in Pichia pastoris and purified using chromatography. Its amino acid sequence is identical to the cationic trypsin derived from porcine pancreas.

Unmodified trypsin may undergo self-cleavage, generating degradation peptides that can interfere with protein sequencing and mass spectrometry peptide mapping, as well as result in reduced enzyme activity during digestion. Furthermore, self-cleavage has been shown to potentially produce chymotrypsin activity.

This product is a recombinantly expressed trypsin, subjected to multiple purification steps to eliminate contaminant enzyme activities. It undergoes reductive methylation, which provides resistance to proteolytic hydrolysis during standalone enzymatic digestion or stepwise/mixed digestion with Lys-C in proteomics and peptide mapping studies, ensuring high activity and stability. Its activity is inhibited by serine protease inhibitors, such as TLCK and PMSF, as well as metal ion chelators like EDTA.

Quality Parameters​ 

• Appearance: White or off-white lyophilized powder
•  Purity: ≥95%
• Molecular Weight: 23kDa ± 2.3kDa 
• Specific Activity: ≥3800 USP/mg 
• Endotoxin: <1 EU/mg 

Advantages

• Excellent Stability: The sequencing-grade trypsin MTP demonstrates exceptional stability. When used at 0.1mg/ml in 50mmol NH₄HCO₃ digestion buffer and incubated overnight (15 hours) at 37°C in a water bath, the enzyme retains over 90% of its activity.
• High Specificity: Using sequencing-grade trypsin MTP at a 1:50 (w/w) ratio to digest the human insulin B-chain, the digestion products were analyzed via RP-HPLC at a wavelength of 214nm. When comparing the digestion products after 16 hours to those after 2 hours, the peptide peak ratio increases, and the number of peaks remains consistent at 2, without further increase over time.

Recommended Usage

• Dissolution Buffer: 10mmol/L HCl or CH₃COOH with 20mmol/L CaCl₂. After dissolution, aliquot and store below -15°C.
• Digestion Buffer: 50mmol/L Tris-HCl or 50mmol/L NH₄HCO₃ with 1mmol/L CaCl₂, pH 7.6–8.0.
• Recommended Conditions: Mix the enzyme with the target protein (concentration: 0.5–1.0mg/ml) at a ratio of 1:20–1:100 in the digestion buffer (50mmol/L Tris-HCl or NH₄HCO₃ with 1mmol/L CaCl₂) and incubate at 30–37°C.

For denatured proteins, it is recommended to dilute urea or guanidine hydrochloride to below 1mol/L before digestion.

Storage Conditions

• Storage Stability: Lyophilized powder remains stable for 24 months when stored below -15°C. After dissolution in HCl or CH₃COOH, aliquot and store frozen below -15°C.
• Transport Stability: Shipped with ice packs to maintain activity stability.