Catalogue number
PRO-585
Synonyms
rTEV, TEV, P1 protease.
Introduction
TEV protease is the common name for the 27 kDa catalytic domain of the Nuclear Inclusion a (NIa) protein encoded by the tobacco etch virus (TEV). Because its sequence specificity is far more stringent than that of factor Xa, thrombin, or enterokinase, TEV protease is a very useful reagent for cleaving fusion proteins. TEV protease recognizes a linear epitope of the general form E-Xaa-Xaa-Y -Xaa-Q-(G/S), with cleavage occurring between Q and G or Q and S. The most commonly used sequence is ENLYFQG.
Description
Recombinant TEV Protease (rTEV) is a site-specific protease purified from E. coli. The protease can be used for the removal of affinity tags from fusion proteins. The seven-amino-acid recognition site for rTEV is Glu-Asn-Leu-Tyr-Phe-Gln-Gly with cleavage occurring between Gln and Gly. The optimal temperature for cleavage is 30°C; however, the enzyme can be used at temperatures as low as 4°C.
The rTEV contains His tag.
The rTEV is purified by proprietary chromatographic techniques.
Source
Escherichia Coli.
Physical Appearance
Sterile liquid formulation.
Formulation
The rTEV contains 25mM Tris, 75mM NaCl, 5mM EDTA, 10mM GSH, 50% Glycerol.
Unit Definition
One unit is defined as the amount of enzyme needed to cleave 3 ug 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.
Stability
rTEV although stable at 4°C for 1 week, should be stored below -18°C.
Please prevent freeze thaw cycles.
Purity
Greater than 90.0% as determined by analysis by SDS-PAGE.
Cleavage Conditions
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 incubations are recommended. Reactions at 4°C will require longer incubation times and/or more rTEV.
Safety Data Sheet
Usage
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
