greenTEV

• Species: Tobaco Etch Virus
• Tags: GFP, N-terminal and His-Tag, C-terminal
• Sequence without tags (AA 5-236):
  KGPRDYNPISSSICHLTNESDGHTTSLYGIGFGPFIITNKHLFRRNNGTLVVQSLHGVFK
  VKDTTTLQQHLVDGRDMIIIRMPKDFPPFPQKLKFREPQREERICLVTTNFQTKSMSSMV
  SDTSCTFPSGDGIFWKHWIQTKDGQCGSPLVSTRDGFIVGIHSASNFTNTNNYFTSVPKN
  FMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEEPFQPVKEATQLMNE
  
  

• Expression Host: E. coli
• Formulation: 
  Liquid: 50 mM Tris, 150 mM NaCl, 0.5 mM EDTA, 40% Glycerol; pH 8.0
  Lyophilized: 50 mM Tris, 150 mM NaCl, 0.5 mM EDTA, 5% Trehalose; pH 8.0
• Format: 
  Liquid: stored at -20 °C and shipped on blue ice
  Lyophilized: stored at -80 °C and shipped at ambient temperature.
  We recommend reconstituting the enzyme in 40 % Glycerol (w/v).
  In case of 0.2 kU, add 10 µl of 40 % Glycerol (w/v)
  In case of 1 kU, add 50 µl of 40 % Glycerol (w/v).
  In case of 10 kU, add 500 µl of 40 % Glycerol (w/v).
  
• Purity: > 85% as determined by SDS-PAGE
• Activity:
  ≥ 20 Units/µl (determined by cleavage of control protein)
  ≥ 0.25 µmol/min/mg (determined by cleavage of labeled peptide (Fluorometric assay), TEV Protease Activity Kit (Abcam))
• Unit definition: One unit of greenTEV will cleave 3 µg of a fusion protein to 98 % in 1 hour 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 represents the catalytic domain of the nuclear inclusion a (NIa) protein with a molecular weight of 27 kDa encoded by the plant virus Tobacco Etch Virus. "green" indicates fusion of the protease to green fluorescent protein (GFP), which leads to increased stability and solubility of TEV protease. Moreover, greenTEV has been optimized by site directed mutagenesis to prevent autocatalytic cleavage. greenTEV is a highly site-specific cysteine protease that recognizes the amino acid sequence Glu-Asn-Leu-Tyr-Phe-Gln-(Gly/Ser) [ENLYFQ(G/S)] and cleaves between the residues Gln and Gly/Ser. The most commonly used recognition sequence is ENLYFQG. In biotechnology, greenTEV is a versatile enzyme to remove affinity tags from recombinant proteins with high specificity and activity over a wide range of pH, ionic strength and temperatures between 4 °C and 30 °C. The optimal temperature for cleavage is 30 °C. It is recommended to improve cleavage efficiency for each fusion protein by varying the amount of recombinant greenTEV, reaction time, or incubation temperature. The great advantage of greenTEV is its facile removal after cleavage reaction by immobilized metal affinity chromatography (IMAC) since it is equipped with a His-tag. Furthermore, the removal of greenTEV can be monitored instantly by detection of fluorescence in solution - this easy, fast and sensitive method omits time-consuming SDS-PAGE or Western blot analysis.
If the green fluorescence of greenTEV is not suitable and you prefer blue fluorescence as readout, check our blueTEV protease.

SDS-PAGE/Coll. Coomassie	Histogram of marked lane in gel picture

Test Cleavage	Description of Test Cleavage

Stability study of lyophilized TEV (blueTEV is shown as an example)

Stability study of lyophilized TEV	Description of Stability study

Download Product Information for greenTEV, liquid

Download Product Information for greenTEV, lyophilized