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 applications, including fully automated workflows and applications that require regulatory compliance.
- Recombinant production
- Engineered to be autolysis resistant – eliminating autolysis digestion artifacts
- High digestion efficiency, no missed cleavages
- Rapid digestion times enable high throughput
- Free of chymotrypsin
- Shelf life of 2 years (dried)
- Reproducible and highly pure. Most preferred protease for Multi-Attribute Method (MAM) analysis
- Ideal for use in PNGase F workflows
Catalog Information
| Catalog # | Concentration | Size |
|---|---|---|
| P8119S | Not Applicable | 100 µg |
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Product Information
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. Lack of digestion artifacts from autolysis or chemotryptic-like activity eliminates the need for manual parsing and enables fully automated reporting. This enzyme is ideal for many proteomics applications and workflows, including those using PNGase F, due to its high purity, autolysis resistance, stability, and rapid digestion times.
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.
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- phi29-XT RCA Kit
- T4 DNA Ligase
- Topoisomerase I (E. coli)
- 9°N™ Reverse Gyrase
- Monarch® Spin Plasmid Miniprep Kit
Product Notes
- The reaction requires ATP and Mg2+ and is further stimulated by spermidine. The supplied Gyrase Reaction Buffer incorporates these essential components within an optimized formulation, thereby facilitating enhanced gyrase activity.
- Proprietary reaction buffer enhances gyrase activity.
References
- Gellert, M et al (1976). PNAS. 73, 3872-3876.
- Higgins, N.P. et al. (1978). PNAS. 75, 1773–1777.
Protocols, Manuals & Usage
Protocols for this product
FAQs & Troubleshooting
FAQs
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.
- Proteases
- Proteome Analysis
- Protein Analysis Tools
- Glycobiology & Proteomics
- Glycoprotein Analysis
Reagents Supplied
The following reagents are supplied with this product:
| NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
|---|---|---|---|---|---|
| P8119S | -20 | ||||
| Trypsin-ultra™, Mass Spectrometry Grade (Recombinant) | P8119SVIAL | -20 | 1 x 100 µg | Not Applicable |
Properties & Usage
Reaction Conditions
Buffer of choice, pH 7-9
Trypsin:substrate ratio from 1:10 to 1:200
Incubate at 37°C
Molecular Weight
Apparent: 22.6 kDa
Advantages and Features
Features
- Recombinant and engineered
- Autolysis resistant
- Fast activity
- Free of chymotrypsin
- Eliminates digestion artifacts from autolysis or chemotryptic-like activity
- Compatible with a broad range of proteomic workflows
Application Features
- Digestion of proteins for proteomic analysis by Mass Spectrometry
- Protein and peptide identification, e.g. Host Cell Proteins (HCP) and low abundance variants
- Enables fully automated and high-throughput workflows
Related Products
Companion Products
- Rapid PNGase F
- Endo S
- PNGase F (Glycerol-free), Recombinant
Product Notes
- Supplied in dry format. Store at -20°C.
- 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
- Compared to animal-derived trypsin, recombinant trypsin delivers higher cleavage specificity and minimal autoproteolysis, reducing artifacts and enabling more accurate MAM analysis of product quality attributes (Pohl T. et al. (2023). European Journal of Pharmaceutics and Biopharmaceutics. https://doi.org/10.1016/j.ejpb.2023.04.024
Protocols, Manuals & Usage
Protocols for this product
- In-Solution Digestion Using Trypsin-ultra™, Mass Spectrometry Grade (Recombinant) (NEB #P8119)
- Rapid Digestion Protocol Using Trypsin-ultra™, Mass Spectrometry Grade (Recombinant) (NEB #P8119)
- Filtration Assisted Sample Preparation (FASP) Using Trypsin-ultra™, Mass Spectrometry Grade (Recombinant) (NEB #P8119)
Tools & Resources
Selection Charts
- Protease Selection Chart
FAQs & Troubleshooting
FAQs
Quality, Safety & Legal
Quality Control Assays
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]
- P8119S_v1
Certificate of Analysis
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]
- P8119S_v1_10319952
Safety Data Sheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
- Trypsin-ultra™, Mass Spectrometry Grade (Recombinant)
Legal and Disclaimers
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.
