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 0.50 M Tris-HCl pH 8.0, 10mM DTT and 5 mM EDTA. 

Unit Definition  One unit of rTEV cleaves >85% of 3 ug control substrate in 1 h at 30°C. 

Activity  10,000 Units/1mg. 

Stability  Store rTEV at -70?C for long term or at -20?C for< 6 months. Please prevent freeze-thaw cycles. 

Purity Greater than 90.0% as determined by(a) Analysis by RP-HPLC.

(b) 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. 

Usage  CHI'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.