blueTEV

€39.00

Price excl. shipping costs excl. VAT. For more information, see our shipping policy

SKU: P2020-138 trenzyme

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Description

blueTEV is an optimized TEV protease (Tobacco Etch Virus Protease) fused to a BFP protein. blueTEV contains a His-Tag to facilitate the removal of the protease from the cleavage reaction after completion of cleavage. The removal of blueTEV can be monitored easily by following the fluorescence - this easy, fast and sensitive method omits time-consuming SDS-PAGE or Western blot analysis. The recognition sequence with the highest catalytic activity is ENLYFQ(G/S).

blueTEV is available in liquid or lyophilized form. The format and size can be selected by using the buttons.


Overview

  • Product Name: blueTEV
  • Catalog No.: P2020-137 lyophilized, P2020-138 liquid
  • RefSeq Links: UniProt: Q0GDU8
  • Synonyms: TEV Protease; TEV; Tobacco Etch Virus nuclear-inclusion-a endopeptidase; rTEV; P1 protease; EC 3.4.22.44

The optimized blueTEV protease and the Cleavage and tag control protein are also available together in the blueTEV Cleavage Kit - order it now!

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Customer Testimonial

“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

Sequence Information

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

Product Information

  • 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.

Background Information

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

SDS-Page blueTEV
blueTEV protease histogram densitometry

 

Additional information blueTEV, lyophilized

SDS-PAGE/Coll. Coomassie

Histogram of marked lane in gel picture

SDS-Page blueTEV
Histogram blueTEV

 

Test Cleavage for blueTEV

Test Cleavage

Description of Test Cleavage

Test Cleavage blueTEV
Description of Test Cleavage blueTEV

 

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

Stability study of lyophilized TEV

Description of Stability study

Stability study of lyophilized TEV
Description Stability study of blueTEV

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