概述
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. with the molecular weight of 27 k Da. 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 GST tag sequence by affinity chromatography.
使用说明
This material is offered by Sangon Biotechk for research, laboratory or further evaluation purposes. NOT FOR HUMAN USE.
技术规格
| Physical Appearance | Clear colorless liquid. |
| Source | Escherichia coli. |
| Recommended Conditions for Cleavage of a Fusion Protein | A number of variables can be changed to optimize the cleavage of any specific protein. The amount of rTEV, the temperature of the incubation, and the time needed for cleavage may be examined. If the protein of interest is heat-labile, then 4 °C incubation |
| Unit Definition | One unit is defined as the amount of enzyme needed to cleave 3 μg 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. |
| Purity | > 90 % by SDS-PAGE analysis. |
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