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TurboTEV Protease
[T-1190]
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Description: TurboTEV Protease contains an enhanced form of a catalytic fragment of the N1a protein of Tobacco etch virus (TEV), a cysteine protease that recognizes the cleavage site of Glu-Asn-Leu-Tyr-Phe-Gln-Gly and cleaves between Gln and Gly. TurboTEV Protease is a restriction grade protease that has a robust activity at 4oC with high specificity and great stability. It does not require any special buffer for its activity and can be used in a buffer most suitable for the target protein. TurboTEV Protease is a 52 kDa protein with both GST and His tags so it can be removed by either Ni-chelating or Glutathione (GSH) resin.
Activity, Specificity, and Stability: TurboTEV Protease has a specific activity of at least 10,000 units/mg, using the conventionally defined activity unit (One unit cleaves >85% of 3 ėg control substrate in 1 h at 30oC). In practice, 1 mg (10,000 units) of TurboTEV Protease cleaves >90% of 100 mg of a control target protein at 4oC in 16 hours. No non-specific cleavage has been observed under the same condition when TurboTEV Protease and the control target protein was mixed at 1:10 ratio. TurboTEV Protease retains >80% activity after storage at room temperature for over 65 hours.
Component: TurboTEV Protease: 2 mg/ml in 25 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM TCEP, and 50% glycerol.
Cleavage Conditions 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 g of target protein in a buffer suitable for the target protein at 4oC 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 g 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 (37oC) can be used to achieve high cleavage efficiency without non-specific cleavage of target proteins.
Removal of TurboTEV Protease after Cleavage: 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.
Bulk Quantity: TurboTEV Protease Readily Available
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$300.00
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Note: All products sold by A.G. Scientific are for Laboratory or Manufacturing purposes only. They are not intended for Human Drug, Food Additive, Clinical or Household use.
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