NumaTEV protease, 6x His-Tag

Numaferm’s TEV protease is an improved variant of the Tobacco Etch Virus (TEV) wildtype enzyme. It harbors various beneficial mutations i.e. at T17, N68 and I77 for increased solubility and stability, and at S219 for auto-proteolysis protection compared to the wildtype enzyme. The recognition site of the Numaferm TEV protease is ENLYFQ/S (or G) and is inserted between your target protein (being located at the C-terminus) and the overhang to be removed (being located at the N-terminus).

Cleavage occurs between Q and S (or G). If your target carries an N-terminal S (or G), just insert ENLYFQ without S (or G) and your target is released native with the correct amino acid at the N-terminus. If your target carries an alternative amino acid, insert ENLYFQS (or G) into your construct. After the cleavage reaction, your target carries an additional S (or G) at its N-terminus.

Order

If you wish to produce your target native and without an N-terminal trace, have a look at our Numacut TEV protease (LINK to the Numacut Product Page). It is an evolved version of TEV protease with broadened substrate tolerance accepting almost all amino acids at the N-terminus of your target.
In which cases this might be important? For example, if …

… the terminus of your target is functionally important.
… the protein has stability or solubility issues with the additional amino acid.
… your R&D/preclinical studies were performed with native proteins, and you do not want to change anything “just” for a more efficient process.
… you do not want to take any risks.
Numaferm’s TEV protease is equipped with an N-terminal 6x His-Tag that paves the way for easy and efficient removal from cleavage reactions by immobilized metal-ion affinity chromatography (IMAC).

Pricing

P084 - 1 1000U: 126 €
P084 - 2 2000U: 206 €
P084 - 3 3000U: 263 €
P084 - 10 10000U: 619 €

*Excl. Shipment Costs (No dry-ice shipment needed due to lyophilized formulation)

Properties

Source: E. coli
Biological Activity: ≥ 10,000 U/mL after reconstitution
Unit Definition: One Unit of NumacutTM TEV protease cleaves 80% of 3 µg control substrate containing a S amino acid residue at P1’ position in 1 h at 30°C and pH 7.5.
Molecular Weight: 28.2 kDa
Formulation buffer: Lyophilized from buffer (20 mM Tris/HCl buffer, 100 mM NaCl, 5 mM DTT, 1 mM EDTA, pH 7.5). No preservatives and carrier-free.
Purity: > 90% by SDS-PAGE analysis.
Storage: Upon receipt, store the Numacut TEV protease at -20°C. The product is stable for at least six months. For application, reconstitute Numacut TEV protease by dissolving it with the provided reconstitution buffer. Reconstituted Numacut TEV protease should be stored at -20°C. Avoid thawing/freezing cycles.

Data Specification Sheet

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Safety Data Sheet

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Application

Upon receipt, the product can be reconstituted by dissolving the protease with the provided buffer. It is recommended to use 10 Units of TEV protease for cleavage of 1-25 µg substrate. Incubate the cleavage reaction at 30°C and use pH values of 7-8 for optimal cleavage efficiency. Usually, 1 hour incubation time is sufficient for complete cleavage, but some substrates may need extended incubation times (3-24h). 50 mM Tris/HCl (pH 8.0), 0.5 mM EDTA and 1 mM DTT containing 1 mg/mL of target protein can be regarded as standard reaction conditions. Avoid > 0.5 M Urea or 0.5 M guanidine hydrochloride, pH values below 4 or above 9, and NaCl concentrations of > 200 mM. It is recommended that the cleavage reaction for each fusion protein is optimized by varying the amount of protease, reaction time and incubation temperature.

Contact to Order

Please fill in the form to place your order.
A member of the team will reach out to you with a quote within 24hrs.
For any questions beforehand, do not hesitate to send an E-Mail to
products@numaferm.com or call us on +49 211 975 32 900.

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