Pierce Trypsin Protease, MS Grade

Thermo Scientific Pierce Trypsin Protease, MS Grade, is highly purified porcine trypsin that has been chemically modified for maximum activity and stability in proteomic applications.

Pierce Trypsin Protease, MS Grade, is a serine protease derived from porcine pancreatic extracts. The enzyme is TPCK-treated to eliminate chymotryptic activity and methylated to improve stability during protein digestion. This mass spectrometry-grade, modified trypsin is then repurified, lyophilized in glass vials and packaged in convenient 5 x 20μg, 5 x 100μg, and 1mg fill sizes.

Highlights:

• Exceptional selectivity – cleaves at the carboxyl side of lysine and arginine residues with greater than 95% specificity
• High purity – no detectable chymotrypsin activity
• Enhanced stability – modified and lyophilized for reduced autolytic activity
• Economical – available in larger fill sizes that are more cost effective

Applications:

• In-gel digestion of proteins from 1-D or 2-D gels
• In-solution tryptic digestion of proteins

Product Details:

Trypsin is a serine protease that specifically cleaves at the carboxyl side of lysine and arginine residues. The selectivity of this enzyme is critical for reproducible protein digestion and mass spectrometry-based protein identification. Since chymotrypsin co-purifies with trypsin derived from natural sources, Pierce Trypsin Protease has been treated with TPCK to eliminate chymotryptic activity, improving digestion specificity. Native trypsin is also subject to autolysis which can reduce enzyme stability and efficiency. To reduce autolytic degradation, Pierce Trypsin Protease is chemically modified by methylation, yielding a highly active and more stable form of the enzyme.

In addition to possessing high specific activity and being resistant to autolytic digestion, Pierce Trypsin Protease can tolerate commonly used partially denaturing conditions, such as 0.1% SDS, 1M urea and 10% acetonitrile. Pierce Trypsin Protease is most active in pH ranges pH 7 to 9 and can be reversibly inactivated at pH < 4. The lyophilized enzyme is also stable for > 1 yr when stored at -20°C.

Excellent digestion performance with Thermo Scientific Pierce Trypsin Protease, MS Grade. Base peak chromatograms of a five-protein mixture sample digested with Pierce Trypsin Protease (top two) and Promega* Trypsin Gold (bottom two). Samples (10μg each) were mixed with trypsin at a 1:50 ratio in a 50mM TEAB buffer (pH ~8) and incubated at 37°C for 30 minutes or 3 hours. Digested sample peptides (0.5μg each) were separated using nanoflow high-pressure liquid chromatography for analysis by a Thermo Scientific Velos Pro mass spectrometer. Click image to see full size.

Comparison of the cleavage selectivity of spectrometry-grade trypsin products. Five-protein mixture samples (10μg) were digested with Thermo Scientific Pierce Trypsin Protease or Promega* Trypsin Gold for 3 hours and analyzed by LC/MS using a Thermo Scientific Velos Pro mass spectrometer. Data was searched using Mascot with "no enzyme" digestion settings. Greater than 95% cleavage selectivity for lysine and arginine (K/R) was observed for Pierce Trypsin Protease.

General References:

1. Biringer, R.G., et al. (2006). Brief Func. Genom. and Prot. 5:144-153.
2. Schlosser, A., et al. (2005). Anal. Chem. 77:5243-5250.
3. Wu, S-L., et al. (2005). J. Proteome Res. 4:1155-1170.

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