NEB offers several proteases with different recognition sites.
Modified Trypsin (TPCK-treated) (NEB #P8101) selectively cleaves peptide bonds C-terminal to lysine and arginine residues (1). Endoproteinase AspN (NEB #P8104) (flavastacin) and Endoproteinase GluC (NEB #P8100) (Staphylococcus aureus Protease V8) selectively cleave peptide bonds N-terminal to aspartic acid residues and C-terminal to glutamic acid, respectively. These proteases are ideal for proteome analysis by mass spectrophotometry. Proteinase K (NEB #P8102) is a non-specific, subtilisin-related serine protease with a very high specific activity. Furin (NEB #P8077) is a ubiquitous subtilisin-like proprotein convertase with a minimal cleavage site requirement of Arg-X-X-Arg'.
We also offer several proteases with unique, specific recognition sites. Enterokinase (NEB #P8070) is a specific protease that cleaves after lysine at its cleavage site Asp-Asp-Asp-Asp-Lys. Factor Xa (NEB #P8010) cleaves after the arginine residue in its preferred cleavage site Ile-(Glu or Asp)-Gly-Arg. Genenase™ I (NEB #P8075) is a variant of subtilisin BPN´ that has been engineered to have increased specificity by substituting amino acids in its active site (2,3). NEB recommends using Genenase I to cleave fusion proteins at the site Pro-Gly-Ala-Ala-His-Tyr. These proteases can be used with the pMAL™ protein fusion and purification system (NEB #E8200) during the purification step.
(1) Northrop, J.H. and Kunitz, M. (1931) Science, 73, 262-263. PMID: 17755302
(2) Carter, P. and Wells, J.A. (1987) Science, 237, 394-399. PMID: 3299704
(3) Carter, P. et al. (1991) Biochemistry, 30, 6142-6148. PMID: 2059622
Genenase™ I is a trademark of Genencor International Inc.
pMAL™ is a trademark of New England Biolabs, Inc.
Protocols for Proteases
- Protocol to purify PCR products in preparation for cloning using Proteinase K (P8107)
- Protocol to cleanup DNA Glucosylation/restriction digest and Proteinase using Proteinase K (P8107)
- General protocol to release nucleic acids prior to capillary or gel electrophoresis using Proteinase K (P8107)
- Reaction conditions for Simultaneous Digestion of IgG with IdeZ Protease (IgG-specific) and PNGase F (fragmentation and deglycosylation) (P0770)
- Reaction Conditions for IdeZ Protease (IgG-specific) (P0770)
- Typical Reaction Conditions for Endoproteinase LysC (P8109)
- In-gel Digestion Protocol for Endoproteinase LysC (P8109)
- Protocol using Trypsin-ultra™, Mass Spectrometry Grade (P8101)
- Typical Trypsin and GluC Co-Digest Reaction Conditions (P8100)
- Typical GluC In-Gel Digest Reaction Conditions (P8100)
- Typical GluC Digest Reaction Conditions (P8100)
- Trypsin Digestion Protocol using NEB Trypsin-ultra™ and the FASP Kit
- Using recombinant Cas9 nuclease to assess locus modification in genome editing experiments (NEB #M0386)
The Glycoproteomics brochure provides information on the suite of endo- and exoglycosidases, and deglycosylation enzymes offered by NEB.
Other Tools & Resources
Protease Selection Chart
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at email@example.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.