Trypsin-ultra™, Mass Spectrometry Grade

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
P8101S     -20       Trypsin-ultra™, Mass Spectrometry Grade P8101SVIAL -20 5 x 20 µg Not Applicable   Trypsin-ultra, Reaction Buffer B8101SVIAL -20 1 x 1.5 ml 2 X

Application Features

• Digestion of proteins for proteomic analysis by Mass Spectrometry
• Protein and peptide identification

Companion Products

• IdeZ Protease (IgG-specific)
• Rapid PNGase F Antibody Standard
• Endo S
• Rapid PNGase F
• Rapid™ PNGase F (non-reducing format)
• Endoproteinase GluC
• Endoproteinase AspN
• Endoproteinase LysC
• α-Lytic Protease

1. Substrate must be in phosphate-free buffer to prevent calcium precipitation with both reconstituted enzyme and enzyme buffer.
2. Trypsin-ultra, Mass Spectrometry Grade is acetylated on multiple lysine residues. This protein appears as a single band on SDS-PAGE. This sequence is also available at www.neb.com.
3. Storage Conditions: Supplied in dry format from a sodium acetate and calcium chloride buffer. Store at -20°C.
   
4. Can be stored frozen in solution at -20°C for up to 2 weeks. A decrease in activity will occur if stored in solution. Use only freshly reconstituted protease for best results.

1. Northrop, J.H. and Kunitz, M. (1931). Isolation of protein crystals possessing tryptic activity. Science. 73,
2. Perona, J.J. and Craik, C.S. (1995). Structural basis of substrate specificity in the serine proteases. Protein Sci.. 4, 337-360.

1. Protocol using Trypsin-ultra™, Mass Spectrometry Grade (P8101)
2. Trypsin Digestion Protocol using NEB Trypsin-ultra™ and the FASP Kit

• AppNote_Unbiased_and_Fast_IgG_Deglycosylation _for_Accurate_N-glycan_Analysis_using_Rapid_PNGase_F
• AppNote_Glycan_Analysis_of_Murine_IgG2a_by_Enzymatic_Digestion_with_PNGase_F_and_Trypsin
• Glycan Analysis of Murine IgG2a by Enzymatic Digestion with PNGase F and Trypsin, Followed by Mass Spectrometric Analysis

• Protease Selection Chart

1. Is there a compatible buffer for digesting with Trypsin (P8101S) and Endoproteinase GluC (P8100S) together?
2. I have a very low concentration of protein and would prefer not to denature as a separate step with buffer exchange before digestion. What denaturants can he use in the Trypsin reaction itself?
3. I would like to use Trypsin to digest some proteins which are present in a cell culture supernatant. The supernatant is composed of DMEM supplemented with 0.2% BSA and antibiotics. Will the enzyme work in the cellular supernatant? The final reaction volume will be 100-300 ul.
4. I am using Trypsin and am wondering about specificity. Does it cut at additional sites when in high concentration?
5. I want to know if calcium can be left out of the buffer since it is causing my DNA-protein complex to precipitate.
6. I want to use Trypsin on 20 ug of protein. How much enzyme do I need to use?
7. How does one do a Trypsin in-gel digest?
8. Is there a simple way to remove Trypsin after protein cleavage?

• Glycoproteomics Brochure
• Glycoproteomics Technical Guide

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.

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]

• P8101S_v1
• P8101S_v2

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]

• P8101S_v1_0071705
• P8101S_v1_0081708
• P8101S_v1_0081710
• P8101S_v1_0081802
• P8101S_v1_10012554
• P8101S_v1_10015652
• P8101S_v1_10019877
• P8101S_v1_10018438
• P8101S_v1_10029463
• P8101S_v1_10030172
• P8101S_v1_10033025
• P8101S_v1_10035579
• P8101S_v1_10044620
• P8101S_v1_10047781
• P8101S_v1_10052776
• P8101S_v1_10059999
• P8101S_v1_10062892
• P8101S_v1_10071484
• P8101S_v1_10075957
• P8101S_v1_10076478
• P8101S_v1_10080304
• P8101S_v1_10087272
• P8101S_v1_10097701
• P8101S_v1_10101728
• P8101S_v1_10108677
• P8101S_v1_10122072
• P8101S_v1_10128173
• P8101S_v2_10129470
• P8101S_v2_10130272
• P8101S_v2_10140352
• P8101S_v2_10149078
• P8101S_v2_10154365
• P8101S_v2_10160715
• P8101S_v2_10169760
• P8101S_v2_10176430
• P8101S_v2_10185557
• P8101S_v2_10191560
• P8101S_v2_10191798
• P8101S_v2_10202227
• P8101S_v2_10206171
• P8101S_v2_10216920
• P8101S_v2_10226036
• P8101S_v2_10247078
• P8101S_v2_10252403
• P8101S_v2_10253404
• P8101S_v2_10255662
• P8101S_v2_10256931

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
• Trypsin-ultra, Reaction Buffer

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.