TEV Protease, Recombinant, His-Tag | Affinity Tag Removal

Amid Biosciences TEV Protease is a 27 kDa engineered form of Tobacco Etch Virus (TEV) protease. The optimum recognition site for TEV protease is the seven-amino-acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) [ENLYFQ(G/S)] with cleavage occurring between the Gln and Gly/Ser residues. The enzyme is highly specific and active for its seven-amino acid sequence with minimal off-target effects which makes it an ideal choice to remove affinity tags from fusion proteins after protein purification. The optimal temperature for cleavage is 30°C; however, it can be used at temperature as low as 4°C. Recombinant TEV protease contains an N-terminal His tag and can be easily removed by immobilized metal affinity chromatography (IMAC) from the cleavage reaction. The enzyme is compatible for both in-solution and on-column cleavage reactions. TEV Protease is purified from E. coli by proprietary chromatographic techniques and is >90% pure when visualized on an SDS-PAGE gel.

TEV Protease is supplied with a vial of 20X TEV buffer (1 M Tris-HCl (pH 8.0), 10 mM EDTA) and a vial of 100 mM DTT.

• Highly specific cleavage activity
• Enhanced enzyme stability for prolonged protease activity
• Activity over a broad temperature (+4°C to 30°C) and pH (6.0 to 8.5) range
• His tag to facilitate its removal from the digested protein samples

Activity: 10,000 U/ml (~10,000 U/mg)

SKU # TVP-301-1 (1000 Units), TVP-301-5 (5000 Units).

Unit Definition

One unit of TEV Protease cleaves ≥85% of 3 µg control substrate in 1 hour at 30°C.

Unit Assay Conditions

The cleavage assay is performed in 1X TEV Buffer (50 mM Tris-HCl, pH 8.0, 0.5 mM EDTA) and 1 mM DTT with 1 unit of enzyme  and 3 µg control substrate at 30°C for 1 hour in a total volume of 30 µl.

Storage: Protease and the supplied components must be stored at -20°C.

International Shipping:  Product requires shipping on ice packs. Please contact info@amidbiosciences.com for shipment estimates

This product is intended for laboratory research use only.

TEV Protease Product Manual

References

 1. Parks et al. Expression and purification of a recombinant tobacco etch virus NIa proteinase: biochemical analyses of the full-length and a naturally occurring truncated proteinase form. Virology, 1995 Jun 20; 210(1): 194-201.

2. Kapust et al. Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Protein Eng., 2001 Dec; 14(12): 993-1000.

3. Cesaratto et al. Tobacco Etch Virus protease: A shortcut across biotechnologies. Journal of Biotechnology, 2016, 231, 239–249.