Additional Information
Product Name: | 6xHis-TEV Protease |
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Also Known As: | TEV protease, NIa, Tobacco etch virus protease |
Catalog No.: | P1001-200 |
Size: | 200 µg |
Molecular Weight: | 27 kDa |
Species: | Tabacco Etch Virus |
Source: | Bacterial recombinant |
Stock: | 100 mM Tris, pH 8.5 at 4 0C, 500 mM NaCl, 5mM DTT, 0.5 mM EDTA and 50% Glycerol |
Concentration: | See tube label |
Quality Assurance: | > 90% purity based on SDS-PAGE. |
Storage: | Store at -80°C; avoid multiple freeze-thaw cycles |
PDF Data Sheet: | PDF Datasheet, MSDS |
NCBI RefSeq: | N/A |
Image(s): | |
Shipping Method: | Dry ice shipping |
References: | 1. Lucast LJ.,et al. (2001) Biotechniques 30, 544. |
Details
Tobacco Etch Virus (TEV) protease is a cysteine protease that is commonly used to remove fusion tags, including His, GST and MBP, at the N or C termini of recombinant proteins. TEV protease specifically recognizes a seven amino acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser (ENLYFQG/S) and cleaves between Gln and Gly/Ser under a wide range of conditions.
Recombinant 6xHis-TEV was expressed in E. Coli and purified to near homogeneity. A Ser219Asn (S219N) substitution was introduced that has no effect on TEV protease activity, but inhibits auto-cleavage and thus, stabilizing TEV protease during incubation and long-term storage.
We recommend using 3% supplied TEV protease (w/w) to preform reactions at 4 0C for 12-16 h or at 30 0C for 1-4 h. Depending on specific substrates, TEV protease concentration, incubation time, and reaction temperature should be optimized. 6xHis-TEV can be removed by Ni-NTA resin after incubation.
TEV protease maintains high activity under a wide range of temperatures (4 0C - 30 0C), pH (6.0 - 8.5) and buffers, and in the presence of commonly used protease inhibitors.