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

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

Description

DTT‑Free TEV Plus Protease (His/Halo‑tag) is a recombinant Tobacco Etch Virus (TEV) cysteine protease expressed in E. coli, engineered with both a 10× His‑tag and a Halo‑tag. This enzyme is DTT‑free, highly specific, highly active, and highly stable, requiring no reducing agents such as DTT, TCEP, or β‑mercaptoethanol to maintain activity. It specifically recognizes the heptapeptide sequence Glu‑Asn‑Leu‑Tyr‑Phe‑Gln‑Gly/Ser and cleaves between the Gln and Gly/Ser residues. TEV protease is widely used to remove fusion tags such as Glutathione S‑transferase (GST), His‑tags, or other affinity tags from recombinant proteins. The application and usage of this product are identical to TEV Plus Protease (His‑tag), and both enzymes recognize and cleave the same peptide sequence.

Through targeted optimization, this product eliminates the need for reducing agents to maintain protease activity. Compared with TEV Plus Protease (His‑tag), the DTT‑Free TEV Plus Protease avoids the risk of disrupting disulfide bonds in target proteins, thereby preserving their native structure and function. In reaction buffers without DTT, this product exhibits enzymatic activity comparable to TEV Plus Protease (His‑tag) in DTT‑containing buffers. In contrast, the activity of TEV Plus Protease (His‑tag) decreases significantly in the absence of DTT.

It is recommended to place fusion tags such as GST, His, or Halo at the N‑terminus of the recombinant protein and insert the TEV protease–specific heptapeptide recognition sequence between the tag and the target protein. After TEV cleavage, only a single extra Gly or Ser residue remains at the N‑terminus of the target protein, thereby minimizing any impact on its structure or biological function.

The optimal cleavage temperature for this product is 30°C, and it maintains high activity within the 29–34°C range. However, its activity decreases sharply at 37°C or above. To better preserve the structure and biological activity of the target protein, we recommend performing overnight digestion at 4°C. This product is active within the pH 6.0–9.0 range but loses activity at pH ≤ 5.

The enzymatic activity of this product is not inhibited by common serine protease inhibitors such as PMSF, AEBSF, Bestatin, Pepstatin, E‑64, TLCK, or EDTA. However, cysteine‑reactive protease inhibitors such as NEM or IAA significantly inhibit the activity of DTT‑Free TEV Plus Protease, because the native enzyme requires Cys151 to maintain catalytic function.

This product carries both a 10× His‑tag and a Halo‑tag, with an approximate molecular weight of 63 kDa and a purity of ≥95%. A version containing only the His‑tag—DTT‑Free TEV Plus Protease (His‑tag)—is also available.

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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 Conditions

Store at –20°C. Stable for two years under recommended conditions.

Notes

• This product is intended solely for use by trained professionals in scientific research. It is not for use in clinical diagnosis or therapy, not for use in food or pharmaceuticals, and must not be stored in residential settings.
• For your safety and health, please wear a lab coat and disposable gloves when handling this product.

Related: 

• TEV Protease
• TEV Protease (High salt tolerance, His tag)
• Enhanced TEV Protease
• TEV Plus Protease (His-tag)
• TEV Plus Protease (GST/His-tag)
• DTT-Free TEV Plus Protease (His-tag)
• DTT-Free TEV Plus Protease (His/Halo-tag)

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

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