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

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

Description

TEV Plus Protease (His-tag) is a cysteine protease from Tobacco Etch Virus (TEV), recombinantly expressed in E. coli and tagged with a 6×His sequence. It exhibits high specificity, strong activity, and excellent stability. TEV Plus Protease specifically recognizes the heptapeptide sequence Glu-Asn-Leu-Tyr-Phe-Gln-Gly/Ser, and cleaves between the Gln and Gly/Ser residues. It is widely used to remove fusion tags such as Glutathione S-transferase (GST), His, or other affinity tags.

Compared to conventional TEV Protease (His-tag), TEV Plus Protease has been optimized for:

• Improved cleavage site specificity
• Higher enzymatic activity
• Enhanced structural stability
• Reduced self-cleavage

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Fusion Protein Design Tip

It is recommended to position GST or His tags at the N-terminus of fusion proteins, and insert the TEV-specific recognition sequence between the tag and the target protein. After cleavage, only one additional Gly/Ser residue remains at the N-terminus of the target protein, minimizing impact on its structure and function.

Optimal Reaction Conditions

• Best cleavage temperature: 30°C
• Active range: 29–34°C
• Activity drops sharply at ≥37°C
• For maximal preservation of protein integrity, overnight cleavage at 4°C is suggested
• Active between pH 6.0–9.0; inactivated at pH ≤5

Imidazole Compatibility

TEV Plus Protease retains high activity in 400 mM imidazole, making it suitable for direct cleavage in freshly purified His-tagged protein solutions. Cleavage can be performed concurrently with dialysis or post-dialysis. After digestion, His-tagged TEV Plus Protease, uncleaved fusion proteins, and released His tags can all be removed via nickel affinity chromatography, leaving the target protein in the flow-through.

Inhibitor Profile

• Not inhibited by serine protease inhibitors such as PMSF, AEBSF, bestatin, pepstatin, E-64, TLCK, and EDTA
• Strongly inhibited by cysteine-reactive reagents like NEM or IAA, as Cys151 is essential for protease activity

Activity Definition

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

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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