HomeProtein ToolsTrypsin-ultra™, Mass Spectrometry Grade (Recombinant)
Trypsin-ultra™, Mass Spectrometry Grade (Recombinant)
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) is a serine protease recombinantly expressed in E. coli, originating from porcine trypsin. It is engineered and further chemically modified via methylation to render it highly resistant to autolysis. This enzyme’s high purity and lack of contaminating enzymes, autolysis resistance, and stability make it ideal for a variety of proteomics workflows, including applications that require regulatory compliance.
Recombinant production
Engineered to be autolysis resistant
Permits rapid digest protocols
Free of chymotrypsin
Shelf life of 2 years (dried)
Reproducible and highly pure. Most preferred protease for Multi-Attribute Method (MAM) analysis
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) is a serine endopeptidase. It selectively cleaves peptide bonds C-terminal to lysine and arginine residues. To minimize autolysis, Trypsin-ultra, Mass Spectrometry Grade (Recombinant) has been engineered and further chemically modified by methylation. Recombinant production of the enzyme ensures it is free of contaminants from animals, including chymotrypsin and other proteolytic activities. This enzyme is ideal for many proteomics applications and workflows, including those using PNGase F, due to its high purity, autolysis resistance and stability.
Product Source
Trypsin sequence from Porcine, mutagenized for enhanced resistance to autoproteolysis and expressed in E. coli.
Specificity
Selectively cleaves peptide bonds C-terminal to lysine and arginine residues.
Figure 1: Trypsin-ultra, Mass Spectrometry Grade (Recombinant) shows superior resistance to autolysis, resulting in cleaner baselines free of autolytic peptides
Samples (9 µg) of Trypsin-ultra, Mass Spectrometry Grade (Recombinant), or a different recombinant Trypsin were incubated in 100 mM Tris-HCl, 10 mM CaCl2 (pH 7.2) for 30 minutes at 37°C. After incubation, samples were injected into a Thermo Scientific® Vanquish™ UHPLC system with UV/Vis detection, coupled to a Thermo Scientific Orbitrap Exploris® 480 mass spectrometer with HCD at 32 NCE setting. Data was analyzed using Protein Metrics® (PMI) Byos® software. The chromatogram shows the UV trace at 214 nm, the arrows indicate autolytic peptides confirmed by MS/MS. The peaks corresponding to the intact Trypsin are marked with an asterisk (*).
Figure 2: Absence of autolytic artifacts with Trypsin-ultra, Mass Spectrometry Grade (Recombinant) streamlines the identification of peaks in NIST reference mAb standard
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) gives clean chromatograms free of artifacts, making it ideal for regulatory compliance workflows. By delivering clean baselines, sample peaks can be identified and quantified automatically with high confidence, without the need of additional annotation or analytical expertise. The Reference Material 8671 NIST mAb (Humanized IgG1) Monoclonal Antibody was digested with Trypsin-ultra, Mass Spectrometry Grade (Recombinant) or a different recombinant Trypsin. Antibody samples (90 µg) were denatured, reduced, and alkylated. After buffer exchange and cleanup they were incubated in 100 mM Tris-HCl, 10 mM CaCl2 (pH 7.2) for 30 minutes at 37°C using a trypsin:protein ratio of 1:10. After digestion, samples were injected into a Thermo Scientific Vanquish UHPLC system with UV/Vis detection, coupled to a Thermo Scientific Orbitrap Exploris 480 mass spectrometer with HCD at 32 NCE setting. Data was analyzed using Protein Metrics (PMI) Byos software. The chromatogram shows the UV trace at 214 nm, with the arrows indicating autolytic peptides confirmed by MS/MS.
Figure 3: Maximum peptide coverage after fast, 30-minute digestion with Trypsin-ultra, Mass Spectrometry Grade (Recombinant)
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) gives high-confidence and maximum peptide coverage during rapid digestion workflows, enabling high-throughput, in-process analysis. The Reference Material 8671 NIST mAb (Humanized IgG1) Monoclonal (90 µg) was denatured, reduced, and alkylated. After cleanup, the antibody was digested with Trypsin-ultra, Mass Spectrometry Grade (Recombinant) using a 1:10 trypsin:protein ratio in 100 mM Tris HCl, 10 mM CaCl2 (pH 7.2) for 30 minutes at 37°C. After digestion, samples were injected into a Thermo Scientific Vanquish UHPLC system with UV/Vis detection, coupled to a Thermo Scientific Orbitrap Exploris 480 mass spectrometer with HCD at 32 NCE setting. Data was analyzed using Protein Metrics (PMI) Byos software.
Specific Activity
The specific activity is measured by the µmol of 7-amino-4-methylcoumarin released per minute per mg of enzyme from Pro-Phe-Arg-7-amido-4-methylcoumarin at 37°C in a total reaction volume of 202.5 µL.
Reconstitution
Trypsin-ultra, Mass Spectrometry Grade (Recombinant) is provided as dried solid and should be reconstituted by the addition of 1 mL of high purity water to make a working solution of 0.1 µg/µL.
This product is related to the following categories:
Once reconstituted, aliquot and store at -20°C for up to 6 months. Avoid repeated freeze/thaw cycles.
Standard conditions are 50 mM Bicarbonate Buffer or Tris HCl buffer, pH 7 to 9, enzyme to protein ratio starting at 1:50 to 1:200, incubate for 3 to 12 hr as needed.
Can be used at high trypsin:protein ratios (for instance 1:10) for fast digestion at 37°C for 30 min with complete digestion and no autolytic peptide artifacts
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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