General description
TEV protease is a highly sequence specific serine protease from Tobacco Etch Virus. Due to its high specificity, TEV protease is popular for cleavage of recombinant fusion proteins. The optimal sequence for TEV protease cleavage is ENLYFQ/S is, however, TEV protease is active on a range of substrates with a consensus sequence of EXLYFΦQ\ϕ where X is any residue, Φ is any large or medium hydrophobic residue and ϕ is any small hydrophobic or polar residue (i.e. glycine, serine, alanine, valine, cysteine). Fusion tag removal in-vitro is the most popular usage of TEV protease.
This biotin-tagged TEV protease is expressed in E.coli. Our Biotin-tagged TEV protease does not carry any purification tag other than biotin and is designed to be used for on-column cleavage of fusion proteins containing a TEV protease recognition sequence. This method specifically cleaves the protein of interest from a column-bound fusion protein, leaving the purification domain or tag bound to the affinity column (e.g. Ni-NTA column) and eluting only the protein of interest.
This method is advantageous to post-elution cleavage for several reasons:
- It eliminates most of the impurities normally associated with affinity purification.
- It allows much gentler elution conditions, with an added flexibility in the composition of the elution buffer. This can help to prevent protein aggregation and inactivation.
After cleavage, the biotinylated TEV protease can be removed with any avidin-conjugated or streptavidin-conjugated beads.
Biotinylation of this product is done enzymatically with no effect on its proteolytic activity. It has no other protein purification tags. The product is supplied at a concentration of =10 U/μl in an aqueous buffer containing 20 mM Tris HCl, pH 7.5, 50 mM Sodium Chloride, 1 mM TCEP, 1 mM EDTA and 50% (V/V) glycerol.
Unit Definition
One unit of TEV protease cleaves >85% of 3 μg of control substrate in one hour at pH 8.0 at 30 °C
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- UPC:
- 51284046
- Condition:
- New
- Availability:
- 3-5 Days
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- MPN:
- SAE0118-5KU
- Temperature Control Device:
- Yes
