Product descriptionThis product is a recombinant sequence-specific protease purified to near homogeneity from E. coli that expresses a gene encoding TEV Protease of Tobacco Etch Virus. TEV Protease recognizes a 7 amino acid sequence (EXXYXQ ↓ G/S) and cleaves the peptide bond between glutamine and glycine or serine (indicated by an arrow ↓).Characteristics- Molecular weight : 28.4 kDa- Reaction temperature : 4 ~ 30℃- Optimal pH range : pH 5.5 ~ 8.5 (optimal pH=7)Applications- Removal of fusion tag from recombinant proteinsQuality control- Purity : >95% on SDS-PAGE- Non-specific protease-free- Endonuclease-free- Exonuclease-free- RNase-freeUnit definition
One unit of TEV Protease is the amount of enzyme required to cleave >85% of 3 μg of substrate protein in 1 hr at 30℃ in the 1X TEV Protease buffer.Storage buffer50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 5 mM DTT, 0.1% TritonX-100, 50% glycerol10X TEV Protease buffer
500 mM Tris-HCl (pH 8.0), 5 mM EDTA, 1 mM DTT
- It is important to confirm cleavage efficiency of TEV Protease with each substrate, since it varies greatly depending on the type of substrates used, incubation temperature, incubation period, and the amount of TEV Protease used. Some substrates, despite the presence of a cleavage site, may not be cleaved if the site is not accessible to the enzyme. This can be overcome by inserting polyglycine, polyhistidine, or a FLAG-tag epitope next to a TEV Protease cleavage site. This allows the recognition sequence to be exposed to the TEV Protease.
- After cleavage reaction, TEV Protease can be efficiently removed with a Ni2+-affinity column due to the presence of a polyhistidine tag at its N-terminus. Prior to a Ni2+-affinity column, EDTA and DDT should be removed. The addition of 10 mM Mg2+ can effectively neutralize EDTA. Generally, substrate proteins bound on the affinity resins in a column requires a larger amounts of TEV Protease than proteins in solution.