Recombinant TEV protease (rTEV Protease) is genetically tailored and expressed in E. coli, incorporating a His tag (6X His tag) to ensure both the retention of the natural TEV enzyme's functional activity and superior stability and specificity across a broad temperature spectrum.

 

 

The rTEV Protease serves as a widely utilized enzymatic tool for the precise removal of tag sequences from fusion proteins. Boasting superior site specificity compared to proteases like EK and SUMO, it exhibits high activity and specificity, making it an ideal choice for your applications.

 

Available in various packaging options, our rTEV Protease comes with its corresponding buffer to ensure optimal performance:

 

 

 

Trust our rTEV Protease for efficient and precise tag removal in your fusion protein applications.

 

Features

The core features of TEV protease lie in its precise recognition of the specific seven-amino acid sequence, EXXYXQ↓(G/S), with the cleavage site precisely positioned between glutamine and glycine/serine. This level of sequence specificity surpasses that of proteases like thrombin, factor Xa, and enterokinase. Notably, TEV protease exhibits remarkable resilience, enabling cleavage within a wide range of pH (6.0-8.5) and temperature (4-30°C), thus allowing for flexible adjustment of reaction conditions tailored to the specific needs of the target protein. Furthermore, the rTEV enzyme achieves its peak activity at pH 7.0 and 30°C.

 

Applications:

TEV protease finds extensive application in the removal of fusion tags from recombinant proteins. A key aspect is the incorporation of 6×His at the N-terminus of the rTEV enzyme. Post-cleavage, the rTEV enzyme can be effortlessly eliminated through Ni2+ affinity chromatography, effectively purifying the target protein.

 

 

Product Properties

 

 

 

Table 1. rTEV cleavage activity at different temperatures

 

 

 

Application examples

 

 

1 Prepare the reaction system as indicated in the table above.

2 Thoroughly mix the contents and incubate at 30°C. Collect samples after 1h, 2h, 4h, and 6h, and place them in separate EP tubes.

3 Add an adequate amount of 5×SDS Loading Buffer to each of the EP tubes and store them at -20°C until the enzyme digestion is complete.

4 Subject the enzyme digestion samples to SDS-PAGE electrophoresis analysis to assess the digestion effect and adjust the required enzyme amount and reaction time accordingly.

5 If the target protein is unstable at 30°C, conduct the reaction overnight at 4°C (approximately 16 hours).

 

 

Transportation and Storage

 

For transportation, keep at low temperature. Upon storage, maintain at -20℃ for a validity period of at least 2 years.

 

Common Factors Affecting the Activity of Natural TEV Enzyme

 

PMSF and AEBSF (1 mM),

TLCK (1 mM),

Bestatin (1 mg/mL), EDTA (1 mM), PestatinA (1 mM), and E-64 (3 mg/mL) - none of these protease inhibitors have any effect on the activity of rTEV.

Other Considerations:

 

Zn2+ concentration above 5 mM can affect the activity of rTEV.

Reagents that react with cysteine are also potential inhibitors of rTEV.

 

 

Common Question

 

Q: What are the advantages of this product?

A: Recombinant TEV protease (rTEV) is a genetically engineered and purified protease. It retains the functionality of the natural TEV enzyme while eliminating its self-enzymatic issues. It exhibits enhanced stability and specificity across a wide temperature range.

 

Q: What experiments can this product be used for?

A: rTEV is a widely used tool enzyme, specifically designed to remove affinity tags from fusion proteins.

 

Q: What is the optimal enzyme digestion time and temperature for this product?
A: 3-3.5h at 16-30℃ is the best.