Performance of cleavage conditions can be easily controlled and visualized by SDS-PAGE using the Cleavage and tag control protein. TEV cleavage results in two cleavage products of 42.8 kDa and 47.7 kDa.
The optimized TEV protease and the Cleavage and tag control protein are also available separately.
greenTEV is an optimized TEV protease (Tobacco Etch Virus Protease) fused to a GFP protein. greenTEV contains an His-Tag to facilitate the removal of the protease from the cleavage reaction after completion of cleavage. The removal of greenTEV can be monitored easily by following the fluorescence. The recognition sequence with the highest catalytic activity is ENLYFQ(G/S).
Product Name: greenTEV & Cleavage and tag control protein
Catalog No.: P2020-CK1 / CK2
- RefSeq Links: UniProt: Q0GDU8
Synonyms: TEV Protease; TEV; Tobacco Etch Virus nuclear-inclusion-a endopeptidase; rTEV; P1 protease; EC 126.96.36.199; control protein, reference protein, cleavage control protein, multiple tag control protein, universal reference protein
“We highly valued the fast and excellent communication and the high flexibility of the team! For any future project, we will preferably entrust in trenzyme’s protein production services.”
Dr. Thore Hettmann
Probiodrug AG, Halle/Saale, Germany
- Species: Tobaco Etch Virus / artificial
greenTEV: GFP, N-terminal and His-Tag, C-terminal
Cleavage and tag control protein: Twin-Strep, HA, T7, Avi, FLAG, and His-tag; N-terminal MBP and C-terminal sfGFP fusion
Sequence greenTEV without tags (AA 5-236):
- Expression Host: E. coli
greenTEV: 50 mM Tris, 150 mM NaCl, 0.5 mM EDTA, 40% Glycerol; pH 8.0
Cleavage and tag control protein: PBS; pH 7.4
Format: Liquid, stored at -20 °C and shipped on blue ice
- Purity: > 85% as determined by SDS-PAGE
- Activity greenTEV: > 0.25 µmol/min/mg (determined by cleavage of labeled peptide (Fluorometric assay), TEV Protease Activity Kit (Abcam))
- Unit definition greenTEV: One unit of greenTEV will cleave 60 µg of a fusion protein to 98 % in 24 hours at room temperature. It is recommended to optimize cleavage conditions for each fusion protein by varying the amount of greenTEV, reaction time, or temperature
greenTEV is an high specific cystein protease (TEV protease, Tobacco Etch Virus Protease) fused to a GFP protein. greenTEV is a highly site-specific cysteine protease optimized for cleavage of tags from fusion proteins containing a TEV-site. The optimal temperature for cleavage is 30°C; also it can be used at temperature as low as 4°C. It is recommended that the cleavage for each fusion protein be optimized by varying the amount of recombinant greenTEV protease, reaction time, or incubation temperature. Since the greenTEV contains an His-Tag, it can easily be removed by Ni2+ affinity resin. The optimum recognition site for this enzyme is the sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) [ENLYFQ(G/S)] and cleavage occurs between the Gln and Gly/Ser residues. The most commonly used sequence is ENLYFQG.
The Cleavage and tag control protein is a versatile protein consisting of various cleavage sites for proteases including TEV, Enterokinase, Thrombin, Factor Xa and PreCission. Protease cleavage results in generation of two fragments that have similar molecular weights, which are as follows: - TEV cleavage: 42.8 kDa and 47.7 kDa - Enterokinase: 43.5 kDa, 20.1 kDa and 26.9 kDa - Thrombin: 43.9 kDa and 46.6 kDa - Factor Xa: 44.3 kDa and 46.2 kDa - PreCission: 45.3 kDa and 45.3 kDa Moreover, the 90.5 kDa fusion protein is equipped with multiple tags such as Twin-Strep-, HA-, FLAG-, and His-tag, for instance, that may serve as positive control performing specific Western blot analyses or any functional assay. Since the protein is fused to GFP, it is additionally applicable as fluorescence control. Thus, the cleavage and tag control protein provides an excellent tool to monitor protease cleavage reactions and specific Western blot analyses as well as fluorescence experiments.
Additional information greenTEV
Histogram of marked lane in gel picture
Description of Test Cleavage