TEV protease encoded by the tobacco etch virus is a catalytic domain of the Nuclear Inclusion a (NIa) protein. It is consists of 241 a.a. amino acids with the molecular weight of 27kDa. TEV recognizes the amino acid sequence of the general form E-X-X-Y-X-Q (or S)/X’, and cleaves between Q (or S)/X’. In this form X and X’ stand for any of the amino acid residues, except that X’ cannot be P. The optimal cleavage site is ENLYFQ/G. As having the absolute specificity and wildly using conditions like broad pH range and ionic strength, the TEV protease became more versatile than EK, thrombin and other protease used in biochemical applications, especially recombinant protein production. The optimal temperature for cleavage is 30°C; however, the enzyme can be used at temperatures as low as 4°C. Following digestion, TEV Protease can be removed from the reaction via the His tag sequence by Ni2+-chelate affinity chromatography.
Source
Escherichia coli.
Molecular Weight
One unit is defined as the amount of enzyme needed to cleave 3 mug of fusion protein in 1 hour to 85 % completion at 30°C in a buffer containing 50 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, and 1 mM DTT.
Biological Activity
Appearance
Clear colorless liquid.
Formulation
A 0.2 um filtered solution in 25 mM Tris-HCl, pH 8.0, 75 mM NaCl, 5 mM EDTA, 10 mM GSH, with 50 % Glycerol.
Endotoxin
Reconstitution
Stability and Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 6 months from date of receipt, -20 to -70 °C as supplied.- 3 months, -20 to -70 °C under sterile conditions after opening.