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 4°C 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 easily removed by either Ni-chelating or Glutathione (GSH) resin along with the cleaved tag. In practice, 1 mg (10,000 units) of TurboTEV Protease cleaves >90% of 100 mg of a control target protein at 4°C 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.
25 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM TCEP, 50% glycerol
Concentration
2 mg/ml
MW
52 kDa
Specific Activity
≥10 units/µg
Unit Definition
One unit cleaves >85% of 3 µg control substrate in 1 hour at 30°C
UniProt #
P04517
References
1. Timmer JC, Salvesen GS. Methods Mol Biol. 2011;753:243-55. 2. Bradley C BC Frew et al. 2012. PLoS Pathogens, 8(4), e1002659.
Storage/Stability
>6 months at –20°C. TurboTEV retains >80% activity after storage at room temperature for over 65 hours.
Assay Conditions
A 49 kDa GST-fusion protein (C) at 1 mg/ml is incubated with TurboTEV or TEV Protease in a buffer of 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, 14 mM β-mercaptoethanol at 4°C for 16 hours. The cleaved products are 27 kDa and 22 kDa. No non-specific cleavage has been observed under the same condition when TurboTEV Protease and the control target protein were mixed at 1:10 ratio.
Instructions for Use
Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.
Applications
Immobilized TEV protease on streptavidin-agarose is an efficient tool for fusion protein cleavage. TEV protease has also been used in a study to investigate proteolytic processing of nlrp1b for inflammasome activity.
Powered by Bioz
