blueTEV

• 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 blueTEV cleaves > 85 % of 3 µg of a fusion protein in 1 hour at room temperature. Cleavage overnight increases cleavage efficiency to > 99 %. It is recommended to optimize cleavage conditions for each protein by varying the amount of blueTEV, reaction time, or temperature.

blueTEV 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. "blue" indicates fusion of the protease to blue fluorescent protein (BFP), which leads to increased stability and solubility of TEV protease. Moreover, blueTEV has been optimized by site directed mutagenesis to prevent autocatalytic cleavage. blueTEV 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, blueTEV 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 blueTEV, reaction time, or incubation temperature. The great advantage of blueTEV 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 blueTEV 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 blue fluorescence of blueTEV is not suitable and you prefer green fluorescence as readout, check our greenTEV protease.

 Additional information blueTEV, liquid

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

Additional information blueTEV, lyophilized

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

Test Cleavage for blueTEV

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 blueTEV, liquid

Download Product Information for blueTEV, lyophilized