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Product Details
Product Details
TEV Protease is a recombinant form of Tobacco Etch Virus protease expressed in Escherichia coli. This protease is used to cleave affinity tags from fusion proteins. The TEV Protease recognition sequence with the highest catalytic efficiency is ENLYFQS/G, the cleavage site is between Q and S/G. TEV protease contains N-terminus His-tag, it can be removed by Ni2+ affinity resin for purification of the target protein.
Materials Provided
ComponentsSizeTEV Protease1000 U/5000 UTEV Protease 10× Reaction Buffer1 mL/5 mLApplication
Removal of affinity purification tags from fusion proteins.
Unit Definition
1 unit of TEV Protease will cleave 2 µg of MBP-fusion protein to 95% completion in a total reaction volume of 10 µl in 1 hour at 30°C in 50 mM Tris-HCl (pH 7.5 @ 25°C) with 0.5 mM EDTA and 1 mM DTT.
Purity
>95% as determined by SDS-PAGE.
>95% as determined by SEC-HPLC.
Enzyme Activity
>1 U/μL
Endotoxin
Less than 1.0 EU per μg by the LAL method / rFC method.
Formulation
Supplied as 0.2 μm filtered solution in 50 mM Tris, 250 mM NaCl, pH7.4 with glycerol as protectant.
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Shipping and Storage
This product is supplied and shipped on dry ice.
This product is stable after storage at:
- The product MUST be stored at -20°C or lower upon receipt;
- -20°C for 3 months under sterile conditions.
- *The 10× Reaction Buffer provided in this kit contains DTT, a sulfhydryl reducing agent essential for maintaining enzyme activity. When employing the 10× Reaction Buffer exposed to prolonged storage or repeated freeze-thaw cycles, fresh 10 mM DTT should be supplemented to the Reaction Buffer to ensure maximal enzymatic activity, as oxidative degradation of DTT progressively compromises its reducing capacity.
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Performance Data
SDS-PAGE
The gel was stained with Coomassie Blue. The purity of the protein is greater than 95% (With Star Ribbon Pre-stained Protein Marker).
SEC-HPLC
The purity of TEV Protease (Cat. No. TEE-V5143) was greater than 95% as determined by SEC-HPLC.
Bioactivity
Two fold dilutions of TEV Protease are incubated with 2 μg MBP5-TEV-paramyosin ΔSal and 1X TEV Protease Reaction Buffer in a 10 µl reaction. The reaction mix is incubated at 30°C for 1 hour. Separation of reaction products are visualized by SDS-PAGE (QC tested).
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