TurboTEV Protease has a specific activity of 10,000 units/mg, using the conventionally defined activity unit (One unit cleaves >85% of 3 µg control substrate in 1 h at 30°C). In practice, 1 mg (10,000 units) of TurboTEV Protease cleaves >90% of 100 mg of a target fusion protein at 4°C in 16 hours. No non-specific cleavage has been observed under the same condition when TurboTEV Protease and target fusion protein was mixed at 1:10 ratio. TurboTEV Protease retains >80% activity after storage at room temperature for over 65 hours

Figure 1
A 49 kDa GST-fusion protein (C) at 1 mg/ml is incubated with TurboTEV or TEV Protease at a ratio of (1) 1:50, (2) 1:100, (3) 1:200 (w/w) in a buffer of 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, 14 mM 2-mercaptoethanol at 4°C for 16 hours. The cleaved products are 27 kDa and 22 kDa. TEV is a competitor TEV product

2 mg/ml in 25 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM TCEP, and 50% glycerol. Store at -20°C. Shipped in gel packs

Purified from E. coli expressing the TurboTEV protease gene.

It is recommended to test TurboTEV Protease cleavage with a protease-to-target protein ratio of 1:100 (w/w) or 1 unit of TurboTEV to 10 ug of target protein in a buffer suitable for the target protein at 4°C overnight, with the target protein concentration at 1-2 mg/ml. In most cases, >90% of target protein is cleaved with a TurboTEV-to-target protein ratio of 1:50 to 1:200 or 1 unit TurboTEV to 5-20 ug of target protein (as shown in Figure 1). The efficiency of cleavage may vary due to the sequences around the cleavage site, the conformation and the solubility of the target protein. Due to its high specificity, more TurboTEV Protease (at 1:10 ratio) or longer cleavage time (over a weekend) at higher temperature (37°C) can be used to achieve high cleavage efficiency without non-specific cleavage of target proteins.

TurboTEV Protease contains both GST and His tags. After cleavage of the target protein, TurboTEV Protease is easily removed along with the tags from the cleavage reaction by affinity chromatography using Ni-chelating resin for His-tagged target protein or GSH resin for GST-tagged target protein

Instruction

 TurboTEV Protease (PDF)