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TEV-protease is a highly site-specific cysteine protease that is found in the Tobacco Etch Virus (TEV). The Genaxon TEV protease comes with an N-terminal His-tag for simple removal from the cleavage reaction by immobilization on metal-affinity resins.
TEV protease recognizes a linear epitope of the general form E-Xaa-Xaa-Y-Xaa-Q-(G/S), with cleavage occuring between Q and G or Q and S. The optimum recognition site for this enzyme is the sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) (ENLYFQ(G/S)). The Genaxxon TEV protease contains a S219V mutation against autoproteolysis (self cleavage) and for increased catalytic activity and efficacy compared to the wild type enzyme. It is mainly used for removing affinity tags from purified peptides or proteins. Affinity tags are often applied to facilitate the expression and purification of recombinant proteins. Whereas many tagged proteins retain their structural integrity and biological activity, others clearly do not. Therefore, whenever possible, it is prudent to remove tags from recombinant proteins.
The tobacco vein mottling virus (TVMV) protease is a close relative of TEV protease with a distinct sequence specificity.
Cleavage reactions with TEV protease are performed generally in 50mM Tris-HCl, pH8.0, 0.5mM EDTA, 1mM DTT for 1 hour at 30°C. Other conditions like lower temperatures and the addition of 1mM TCEP in place of DTT can be applied. If different conditions have to be used, the amount of TEV protease added, the incubation time and/or the incubation temperature have to be optimized.
The optimum salt concentration is 0mM but an increasing salt concentration up to 200mM NaCl will only result in a three fold decrease in activity.
- Highly specific - high activity - more specific than Faktor Xa, Thrombin or Enterokinase
Purity: >95% (SDS-PAGE) delivered in 50% glycerol
MW = 28.6 kDa.
Specific Activity: One unit of TEV Protease cleaves >85% of 3μg of control substrate in 1 hour at 30°C.
Applikation:- Protein cleavage - Tag removal from recombinant proteins or peptides
Sicherheits Hinweise / Safety
Klassifizierungen / Classificationeclass-Nr: 32-16-04-10
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