Numacut TEV Protease,
6x His-Tag

The Numacut TEV protease is an improved variant of the Tobacco Etch Virus (TEV) protease with widened substrate tolerance. In comparison to the wild type enzyme, Numacut harbours various advantageous mutations. Primarily, instead of accepting the recognition sequence ENLYFQS (or G) and cleaving between Q and S/G (called P1' position), Numacut can be applied for all amino acids at P1', expect proline (P). In addition, further beneficial mutations increase the specific activity, solubility and stability outperforming the wildtype enzyme significantly. The N-terminal 6x His-Tag paves the way for easy and efficient removal of the Numacut from cleavage reactions by immobilized metal-ion affinity chromatography (IMAC). If you desire your candidate to be sequence-identical without traces, choose Numacut.
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Properties

  • Source: E. coli
  • Biological Activity: > 20,000 U/mL
  • Unit Definition: One Unit of Numacut TEV protease cleaves 80% of 3 µg control substrate in 1 hour at 30°C and pH 8.0.
  • Molecular Weight: 28.2 kDa
  • Formulation: Lyophilized from buffer (20 mM Tris/HCl buffer, 100 mM NaCl, 5 mM DTT, 1 mM EDTA, pH 7.5). No preservatives and carrier-free.
  • Purity: > 95% by SDS-PAGE analyses
  • Storage: Upon receipt, store the Numacut protease at -20°C or reconstitute it by dissolving it with the provided reconstitution buffer. Upon reconstitution, the product is stable for at least six months at -20°C.
Data Specification Sheet
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Safety Data Sheet
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Application

10 Units of Numacut protease are recommended for the cleavage of 1-25 µg substrate (≥ 1 mg/mL) in reaction buffer (50 mM Tris/HCl (pH 8.0), 0.5 mM EDTA and 1 mM DTT). Incubate the cleavage reaction between 30-37°C and at pH 7-8. Usually, a reaction time of 1h is sufficient for quantitative cleavage. Increase the reaction time (≤ 24h) to improve the cleavage efficiency. Avoid > 0.5 M urea or 0.5 M GuHCl, pH values below 6 and above 9, and high salt concentrations ( > 150 mM). Optimization of the cleavage reaction can be achieved by varying the amount of protease, reaction time and incubation temperature.

Case Study - NumaTEV Protease

Digestion of Numaswitch tagged fusion protein

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Please get in touch with us to place your order.
A member of the team will reach out to you to discuss the details of your order.
For any questions beforehand, do not hesitate to send an E-Mail to
info@numaferm.com or call us on +49 211 975 32 900.

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