AcTEV™ Protease specifically recognizes a seven amino acid sequence (Glu-Asn-Leu-Tyr-Phe-Gln-Gly, cleaving between Gln and Gly), making it useful for removing affinity tags from fusion proteins. AcTEV™ Protease is an improved version of Tobacco Etch Virus (TEV) protease that is highly site-specific, highly active, and significantly more stable than native TEV protease, resulting in enhanced long-term activity. AcTEV™ Protease features:

• Highly specific cleavage activity
• Enhanced enzyme stability for prolonged protease activity (see figure)
• Activity over a broad temperature (+4°C to 30°C) and pH (6.0 to 8.5) range
• Six-histidine sequence to facilitate its removal from the digested protein sample
• Greater than 85% single-band purity with no nonspecific protease contamination

Applications
Incubation with AcTEV™ Protease releases the protein of interest from the fusion tag. This is an effective way to remove solubility, secretion, detection, and purification tags from recombinant proteins.

Enzyme specifications
Purified from E. coli expressing the AcTEV™ Protease gene.

Unit definition
One unit of AcTEV™ Protease cleaves 85% of a 3 μg control substrate in 1 hr at 30°C.

Unit reaction conditions
50 mM Tris-HCl (pH 8.0), 0.5 mM EDTA, 1 mM DTT, 3 μg control substrate, and 1 unit enzyme in 30 μL for 1 hr at 30°C. AcTEV™ Protease is functionally tested for the absence of any nonspecific protease activity.