Recombinant TEV Protease (His-tag), 5000 U

Product Information

Product Description/Introduction

Recombinant TEV Protease is recombinant expressed as a His-tag Tobacco Etch Virus (TEV) cysteine protease in Escherichia coli. TEV Protease is a commonly used cysteine protease, which can recognize the seven-amino-acid sequence Glu-Asn-Leu-Tyr-Ph-Gln-Gly /Ser(E-N-L-Y-F-Q-G/S) and cleaves between Gln(Q) and Gly/Ser(G/S) amino acid residues with high specificity. The optimal temperature of Recombinant TEV Protease for cleavage is 30, and the enzyme is active over the range of temperature ( 29-34). However, the enzyme activity will decrease sharply when the temperature reaches or exceeds 37, In the actual process, in order to preserve the structure and biological activity of the target protein as much as possible, it is recommended to use TEV Protease at 4 for overnight digestion. Recombinant TEV Protease is active in the range of pH 6.0 to 9.0, but it will be inactived when the pH is at or less than 5. Another prominent advantage of TEV Protease is that 400 mM imidazoles still have high activity. Therefore, for the purification of target protein containing His tag by nickel column, TEV Protease can be directly added into the purified target protein solution containing high concentration of imidazole, and the imidazoles are removed by dialysis at 4 and digested simultaneously. It can also be digested with TEV Protease after dialysis to remove His tags. Both the His-tagged TEV Protease and the excised His tag in the target protein solution after enzymatic digestion can be removed by binding to the nickel column.

Application: It is often used to remove MBP, GST, His or other tag proteins from fusion proteins. The Recombinant TEV Protease itself has a His tag that binds to the nickel column.

Source: Tobacco Etch Virus (TEV) cysteine protease, recombinant expressed in Escherichia coli, containing His tag.

Enzyme activity: It is defined as one active unit for cleaving more than 85% of 3 μg substrate (protein containing Recombinant TEV Protease cleavage site) within 1 h at 30, pH 8.0.

Inactivation or inhibition: Common serine Protease inhibitors such as PMSF, AEBSF, bestatin, pepstatin, and EDTA do not inhibit TEV Protease activity. However, Protease inhibitors targeting cysteine residues such as NEM or IAA can significantly inhibit the activity of TEV Protease. TEV Protease remained a high activity in higher concentration of imidazole.

Purity and concentration: Purity 95% by SDS-PAGE, 10 U/μL.

Recombinant TEV protease storage solution：50 mM Tris-HCl，250 mM NaCl，1 mM EDTA，5 mM DTT，50% (v/v) Glycerol，pH 8.0。

10×TEV Buffer：500 mM Tris-HCl，500 mM NaCl，5 mM EDTA，10 mM DTT，pH 8.0。

Storage and Shipping Conditions

Ship with wet ice; Store at -20; Valid for 12 months.

Product Components

Assay Protocol/Procedures

1.  Protease digestion system recommended：

2. The reaction system is placed at 30 for 1-4 h. If the target protein is not stable at 30,,the reaction system can be placed at 4 overnight for enzyme digestion (about 16 h)；

3.  10 μL of enzyme digestion product was taken for SDS-PAGE electrophoresis to evaluate the amount of enzyme digestion and reaction conditions.

Note

1. Enzyme products should be placed in an ice box or ice bath when used and stored separately at -20 immediately after use；

2.  For your safety and health, please wear safety glasses, gloves, or protective clothing.

Examples of enzyme digestion applications

Figure 1: Effect of Recombinant TEV Protease digestion of control substrate protein (A+B fusion protein containing Recombinant TEV Protease Enzyme cutting site) (The enzyme digestion conditions were 30 and pH 8.0, respectively for 0 min, 10 min, 20 min, 30 min, 1 h and 2 h, corresponding to Lane 1-6 in turn, and the mass ratio of substrate to enzyme added in the enzyme digestion system was 20:1).

For Research Use Only!

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