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).
Sequence without tags (AA 5-236): KGPRDYNPISSSICHLTNESDGHTTSLYGIGFGPFIITNKHLFRRNNGTLVVQSLHGVFK VKDTTTLQQHLVDGRDMIIIRMPKDFPPFPQKLKFREPQREERICLVTTNFQTKSMSSMV SDTSCTFPSGDGIFWKHWIQTKDGQCGSPLVSTRDGFIVGIHSASNFTNTNNYFTSVPKN FMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEEPFQPVKEATQLMNE
Expression Host:E. coli
Formulation: 50 mM Tris, 150 mM NaCl, 0.5 mM EDTA, 40% Glycerol; pH 8.0
Format: Liquid, stored at -20 °C and shipped on blue ice
Purity: > 85% as determined by SDS-PAGE
Activity: > 0.25 µmol/min/mg (determined by cleavage of Fluorescein-based peptide, TEV Protease Activity Kit (Abcam))
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.