AcTEV™ protease specifically recognizes a seven-amino acid sequence (Glu-Asn-Leu-Tyr-Phe-Gln-Gly, cleaves between Gln and Gly), allowing it to be used to remove the affinity tag from the fusion protein. AcTEV™ Protease is an improved version of the Tobacco Etch Virus (TEV) protease that is highly site-specific, highly active and significantly more stable than the native TEV protease, resulting in enhanced long-term activity. Features of AcTEV™ Protease:

• Highly specific cleavage activity
• Enhanced enzyme stability, thereby enhancing long-term protease activity (see figure)
• Over a wide temperature range (+4°C to 30°C) and pH range (6.0 to 8.5)
•Identifies six histidine sequences, helping to remove them from digested protein samples
•Single band purity is greater than 85% , no non-specific protease contamination

Applications
Incubation with AcTEV™ protease releases the target protein from the fusion tag. This is an efficient method 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 3 µg of control substrate in 1 hour 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/30 µL , 1 h at 30°C. AcTEV™ protease is functionally tested and does not exhibit any non-specific protease activity.