Tobacco Etch Virus Proteinase, mutant, Recombinant, His-tag (mTEVP, TEVP)

Tobacco Etch Virus Proteinase, mutant (mTEVP, Recombinant) is used as a valuable tool for cleaving affinity tags and fusion proteins in recombinant protein purification protocols. The wild type TEVP contains a self-cleavage site that the protein recognizes and cleaves. Since the protein cleaves itself, it is inherently unstable. A mutant TEVP has been generated and tested to have reduced self-cleavage function with no reduction in its target cleavage activity. The protein was expressed in bacteria and purified by Ni affinity chromatography.\n\nApplication:\nSuitable for use in cleaving 6X His-tags from target proteins. After digestion, the cleaved tag and the mTEVP, since they both have His-tags, can be removed by adsorption onto a Ni column. Can also be used to remove Protein A tags.\n\nRecognition Sequence:\nE-N-L-Y-F-Q*G-M------\n*-cleavage point\n\nPurity:\nPurified by nickel chromotography ((same/more than) 90%). Single band (SDS-PAGE) of ~28kD\n\nActivity:\nmTEVP has a cleavage activity of ~8 units/ul\n\nUnit Definition:\nOne unit of mTEVP cleaves (same/more than) 95% of 3ug of control substrate in 1hour at 30 degrees C.\n\nRecommended reaction buffer:\nTEV enzyme assay is carried out in 50mM Tris buffer, pH 8.0, 0.5mM EDTA, 1mM DTT.\n\nOptimal Reaction Temp: \n30 degrees C (minimal temp is not tested)\n\nreaction buffer:\n50 mM Tris buffer, pH 8.0, 0.5 mM EDTA and 1 mM DTT.\n\nStorage Buffer:\n100mM Tris, pH 8.5, 500mM sodium chloride, 50% glycerol, 5mm DTT, 0.5mM EDTA\n\nStorage and Stability:\nMay be stored at 4 degrees C for at least 6 months. For long-term storage, aliquot and store at -70 degrees C. Aliquots are stable for 12 months at -70 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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