Cysteine protease from Tobacco Etch Virus for the removal of affinity tags from fusion proteins under target protein friendly conditions. Genetically engineered for resistance against autolysis, as well as for improved activity and performance. TEV is a cysteine protease that specifically recognizes and cleaves a linear epitope with general sequence E-X-X-Y-X-Q-(G/S) (where X is any amino acid). The cleavage occurs between Q and G/S. The most common sequence is ENLYFQG or ENLYFQS. Resistant to many widely used serine and cysteine protease inhibitors. Robust enzyme active in the wide range of different buffers (with NaCl varied from 0 to 0.4 M and in pH from 4 to 9, enzyme tolerates MES, acetate, phosphate, glycerol and sorbitol). Active in a broad temperature range from 4 to 30°C (the enzyme is 3 times less active at 4°C than at 30°C).
Optimal Reaction Conditions (end conc.):
- 50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA, 1 mM DTT
- cleavage Temperature: 1-6 h at 30°C or 16-24 h at 4-8°C
- Add 10U TEV protease for aprox. 20 µg fusion protein
50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA, 1 mM DTT, 50% (v/v) glycerol.
Store at -20°C (six months) or -80°C (years). No autolysis.
Unit Definition: One unit is the amount of enzyme required to cleave >85% of 3 µg of control substrate (35 kDa fusion protein) in 1 hour at 30°C.
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Manufacturer: BLIRT S.A.