The Science Behind TEV Protease His Tag: Applications, Protocols, and Optimization Tips

In the world of protein engineering and molecular biology, precision tools are vital for achieving high-yield, high-purity results. One such indispensable tool is TEV protease with His tag—a highly specific enzyme used to cleave fusion proteins at defined sites. Backed by innovation and reliability, Gene Universal offers a premium-grade TEV protease designed to optimize purification workflows in both research and biomanufacturing.

As a pioneering technology and service provider in life science research, Gene Universal is committed to advancing global R&D through efficient, cost-effective, and sustainable tools that contribute to the development of biologics and beyond.

What Is TEV Protease His Tag?

TEV (Tobacco Etch Virus) Protease is a recombinant enzyme derived from the Nla protein of the TEV virus. It recognizes a highly specific seven-amino-acid sequence (typically ENLYFQ↓G or S)and cleaves precisely between the glutamine (Q) and glycine (G)/serine (S). The high specificity of TEV protease makes it ideal for removing affinity or solubility tags without damaging the protein of interest.

Gene Universal’s TEV protease is produced via E. coli expression systems and purified through affinity chromatography, incorporating a His tag that allows for its easy removal post-reaction using nickel resin.

Applications in Protein Engineering

The TEV protease His tag system plays a crucial role in various protein expression and purification applications:

  Cleavage of fusion tags to obtain native, untagged proteins for structural or functional studies.

  Improving solubility of recombinant proteins by removing bulky affinity tags post-purification.

  Enhancing protein stability or activity by eliminating extraneous peptide sequences.

  Streamlining downstream processing in therapeutic protein and vaccine production.

Protocol: How to Use TEV Protease His Tag Effectively

To ensure optimal results, follow this step-by-step guide when using Gene Universal’s TEV protease:

✅ Reaction Setup

Prepare the following digestion system in an EP (Eppendorf) tube:

  Fusion Protein: 1 mg

  TEV Protease:10 units (U)

Mix gently to ensure full integration of the components.

✅ Incubation

  Recommended conditions:

      Temperature: 4°C (overnight incubation) for gentle digestion

      Alternative range: 4–30°C, depending on protein stability

      pH: 5.5–8.5 (buffer-dependent)

TEV protease remains active over a wide pH and temperature range, giving users the flexibility to match digestion to their protein's stability profile.

✅ Post-Digestion Analysis

  After the enzymatic reaction,analyze a sample using SDS-PAGE to confirm successful cleavage.

  If you need to remove the TEV protease post-digestion, leverage the His tag by using histidine-tag affinity purification resin (such as Ni-NTA agarose).

Storage Guidelines

Proper storage ensures long-term activity of the enzyme:

  -80°C for long-term storage (up to 2 years)

  -20°C for short-term storage (up to 6 months)

For optimal use, consider aliquoting into small volumes upon first thaw.

Optimization Tips

To fine-tune your TEV protease reactions:

  Ratio Matters: Use a ratio of 1:100 (w/w) of TEV protease to target protein, but adjust based on your experimental needs.

  Buffer Compatibility: Ensure the digestion buffer is compatible with both the protease and your protein.

  Minimize Denaturants: Avoid urea or high concentrations of detergents during the reaction unless proven compatible.

  Time vs. Temperature: Higher temperatures may accelerate cleavage but risk degrading sensitive proteins; 4°C overnight is the safest option.

Why Choose Gene Universal’s TEV Protease?

  High purity recombinant enzyme with reliable performance

  Easy post-reaction removal via His tag affinity

  Flexible and scalable, suitable for both small research labs and industrial production

  Backed by Gene Universal’s commitment to scientific innovation and sustainability

Conclusion

Whether you're conducting small-scale protein research or scaling up biomanufacturing workflows, Gene Universal’s TEV Protease His Tag solution offers precision, convenience, and confidence. With detailed protocols, strong technical support, and proven enzyme activity, it’s a smart choice for protein engineers who demand high-quality tools.

Empower your experiments with Gene Universal—where technology meets trust in life science innovation.