SelecTEV™ Protease is a 27 kD improved form of Tobacco Etch Virus (TEV) protease that has been engineered to be more stable, active, and specific than the native protease. SelecTEV™ Protease is a cysteine protease that recognizes the TEV recognition sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly and cleaves between Gln and Gly. This seven-amino-acid sequence is rarely found in proteins, making SelecTEV™ Protease an ideal choice for tag removal of fusion proteins. The optimal temperature for cleavage is 30°C, however the enzyme is active over a wide temperature range (4 – 30°C) and pH (6.0 – 8.5) to accommodate your specific protein. Following digestion, SelecTEV™ Protease is easily removed from the cleavage reaction by affinity chromatography using the polyhistidine tag at the N-terminus of the protease. SelecTEV™ Protease is purified from E. coli by affinity chromatography using the polyhistidine tag and is 90% pure when visualized on an SDS-PAGE gel.
SelecTEV Protease can be used with the Expresso Solubility and Expression Screening System vectors, which contain a TEV cleavage recognition site.
Coomassie-stained SDS-PAGE gel of TEV cleavage reaction. 10 U of SelecTEV Protease was incubated with 30 µg of fusion protein substrate at 30°C for 1 hour in a 100 µL reaction. Marker (lane M), SelecTEV Protease (Lane 1), Uncleaved fusion substrate (Lane 2), Cleaved substrate and SelecTEV Protease. (Lane 3)
One unit of SelecTEV™ Protease cleaves ≥85% of 3 µg control substrate in 1 hour at 30°C as seen on an SDS-PAGE gel.
Unit Assay Conditions
The cleavage assay is performed in 1X SelecTEV™ Buffer (50 mM Tris-HCl, pH 8.0, 0.5 mM EDTA) and 1 mM DTT with 10 units SelecTEV™Protease and 30 µg control substrate at 30°C for 1 hour in a total volume of 100 µL.