TurboTEV Protease contains an enhanced form of a catalytic fragment of the N1a protein of Tobacco etch virus (TEV), a cysteine protease that recognizes the cleavage site of Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser and cleaves between Gln and Gly/Ser. TurboTEV Protease is a restriction grade protease that has a robust activity at 4°C with high specificity and great stability. It does not require any special buffer for its activity and can be used in a buffer most suitable for the target protein. TurboTEV Protease is a 52 kDa protein with both GST and His tags so it can be easily removed by either Ni-chelating or Glutathione (GSH) resin along with the cleaved tag.
TurboTEV Protease has a specific activity of 10,000 units/mg, using the conventionally defined activity unit (One unit cleaves >85% of 3 µg control substrate in 1 h at 30°C). In practice, 1 mg (10,000 units) of TurboTEV Protease cleaves >90% of 100 mg of a target fusion protein at 4°C in 16 hours. No non-specific cleavage has been observed under the same condition when TurboTEV Protease and target fusion protein was mixed at 1:10 ratio. TurboTEV Protease retains >80% activity after storage at room temperature for over 65 hours
A 49 kDa GST-fusion protein (C) at 1 mg/ml is incubated with TurboTEV or TEV Protease at a ratio of (1) 1:50, (2) 1:100, (3) 1:200 (w/w) in a buffer of 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, 14 mM 2-mercaptoethanol at 4°C for 16 hours. The cleaved products are 27 kDa and 22 kDa. TEV is a competitor TEV product