TEV Protease

Description

TEV Protease (Tobacco etch virus protease) is a recombinant protease derived from tobacco etch virus (TEV) Nla, which is used to excise affinity tags of purified fusion proteins. TEV Protease has strong site specificity and can recognize the seven amino acid sequence (Glu-Asn-Leu-Tyr-Phe-Gln-Gly) of EXXYXQ (G/S), most commonly ENLYFQG, whose cleavage site is between glutamine and glycine or serine. The enzyme is active in a wide range of pH 5.5-8.5 and 4-30 °C, so that the reaction conditions can be modified according to the target proteins. TEV Protease is expresseed and purified from E. coli. TEV Protease has a polyhistidine tag and can be removed by affinity chromatography after the enzyme digestion reaction.

Unite definition

In 1×TEV Buffer (50 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 1 mM DTT), the amount of enzyme required to cleave >85% of the 3 µg control substrate at 30 °C for 1 h is defined as an active unit.

Storage

TEV Protease can be stored for 2 years at -80 °C for a long time or for 6 months at -20 °C after sub-packaged to avoid repeated freezing and thawing. 10 x TEV Buffer can be stored at -20 °C.

Protocol

If the protein is thermally unstable, incubate at 4 °C for a longer time or increase the amount of enzyme used.

1. Set up the following reaction system in EP tube:

2. After mixing the system, incubate at 30 °C. Take 30 μl solution at 1, 2, 4 and 6 h respectively, and place it in a separate EP tube.

3. Add 20 μl 2 x SDS Loading Buffer to the above EP tube and place it at -20 °C.

4. Take 30 μl sample for SDA-PAGE analysis.

Note

To achieve the best digestion result, please ensure that the recombinant protein is partially or completely purified.

Only for research and not intended for treatment of humans or animals