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Cat. No.: TEVPG-1k (for 1,000U)

Cat. No.: TEVPG-10k (for 10,000U)

Description

TEV Plus Protease (GST/His-tag) is a highly specific, active, and stable cysteine protease from Tobacco Etch Virus (TEV), recombinantly expressed in E. coli with both GST and 6×His tags. It specifically recognizes the heptapeptide sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser and cleaves between the Gln and Gly/Ser residues. This enzyme is commonly used to remove fusion tags such as GST, His, or other affinity labels.

The TEV Plus Protease (GST/His-tag) has been engineered for:

• Enhanced site-specific recognition
• Higher enzymatic activity
• Improved stability
• Reduced autolysis

Tag Design Recommendation

For optimal results, design fusion proteins with GST or His tags at the N-terminus, followed by the TEV-specific recognition sequence. After cleavage, the N-terminus of the target protein retains only a Gly/Ser residue, minimizing structural and functional impact.

Optimal Conditions

• Temperature: Best at 30ºC; highly active from 29–34ºC, but activity drops sharply at ≥37ºC
• Suggested protocol: Overnight cleavage at 4ºC for better preservation of protein integrity
• pH range: Active from pH 6.0 to 9.0; loses activity at pH ≤ 5

Imidazole Compatibility

This protease retains high activity in 400 mM imidazole, making it suitable for use directly in freshly purified His-tagged protein solutions. Cleavage can be carried out during dialysis to remove imidazole or afterward. Post-digestion, His-tagged TEV Plus Protease, residual fusion proteins, and tag fragments can be removed via nickel column, leaving the desired protein in the flow-through.

Dual Purification Advantage

This protease is also suitable for use with GST-tagged fusion proteins. After cleavage, incomplete digestion products, free GST tags, and TEV Plus Protease (GST/His-tag) can be removed using GST purification media, with the target protein collected in the flow-through. Compared to the His-tag version, this dual-tag format provides greater flexibility for purification workflows.

Inhibitor

• Not inhibited by common serine protease inhibitors: PMSF, AEBSF, bestatin, pepstatin, E-64, TLCK, EDTA
• Significantly inhibited by cysteine-targeting inhibitors such as NEM or IAA, due to the essential Cys151 residue

Unit Definition

One unit is defined as the amount of enzyme required to cleave ≥85% of 3 µg control substrate in 1 hour at 30ºC, pH 8.0

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Storage

Store at -20ºC. Valid for two years.

Precautions

• This product is intended solely for scientific research by trained professionals. It must not be used for clinical diagnosis or treatment, nor for food or pharmaceutical applications. Do not store in residential areas.
• For your safety and health, wear a lab coat and disposable gloves during use.

Related: 

• TEV Protease
• Enhanced TEV Protease
• TEV Plus Protease (His-tag)
• TEV Plus Protease (GST/His-tag)

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

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