Recombinant TEV Protease Protein, CF

Images

 
There are currently no images for TEV Protease (4469-TP).

Every product we sell is backed by Novus' 100% Guarantee. If you have used this product, please submit your images and reviews to earn reward points.

Product Details

Summary
Reactivity VSpecies Glossary
Applications Enzyme Activity
Format
Carrier-Free

Order Details

Recombinant TEV Protease Protein, CF Summary

Details of Functionality
Measured by its ability to cleave a fusion protein containing the recognition sequence Glu-Asn-Leu-Tyr-Phe-Gln, with the cleavage point after Gln. TEV Protease cleaves ≥50% of the control substrate, as measured under the described conditions. It is recommended that the cleavage for each fusion protein be optimized by varying the amount of Recombinant Viral TEV Protease, reaction time, or incubation temperature.
Source
E. coli-derived viral TEV Protease protein
Ala TEV Protease
Glu2039-Gln2279 (Ser2256Asn)
Accession # NP_062908
Leu 6-His tag
N-terminus C-terminus
Accession #
N-terminal Sequence
Ala
Protein/Peptide Type
Recombinant Enzymes
Purity
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Note
<1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

Theoretical MW
28.5 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
SDS-PAGE
25-30 kDa, reducing conditions
Publications
Read Publication using
4469-TP in the following applications:

Packaging, Storage & Formulations

Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.
Buffer
Supplied as a 0.2 μm filtered solution in Tris, NaCl, EDTA, DTT and Glycerol.
Purity
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Assay Procedure
  • Assay Buffer: 50 mM Tris, 0.5 mM EDTA, 1 mM DTT, pH 8.0
  • Recombinant Viral TEV Protease (rvTEV) (Catalog # 4469-TP)
  • Substrate: Any fusion protein containing the recognition sequence ENLYFQ. The cleavage site is after glutamine (Q).
  • SDS-PAGE followed by protein staining
  1. Dilute rvTEV Protease to 0.02 mg/mL in Assay Buffer.
  2. Dilute Substrate to 0.5 mg/mL in Assay Buffer.
  3. Form reaction mixture by combining 20 µL of diluted Substrate and 20 µL Assay Buffer. Also prepare two additional mixtures, to be used as controls, containing 20 µL of diluted Substrate and 30 µL of Assay Buffer.
  4. Incubate all three mixtures (temperature equilibration), as well as the diluted rvTEV Protease, for 15 minutes at room temperature.
  5. Add 10 µL diluted rvTEV Protease to the reaction mixture, excluding the controls, for a final volume of 50 µL in each reaction.
  6. Incubate the reaction containing the diluted rvTEV Protease as well as one of the controls for 1 hour at room temperature. Keep the other control on ice.
  7. Stop the reactions by mixing equal volumes of reaction mixture (including controls) and 2X reducing SDS-PAGE sample buffer together.  Heat for 5 minutes at 100 °C.
  8. Analyze the cleavage by SDS-PAGE followed by protein staining.
Per Reaction:
  • rvTEV Protease: 0.2 µg
  • Substrate: 10 µg

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant TEV Protease Protein, CF

  • NIa
  • P1 Protease
  • TEV Protease
  • Tobacco Etch Virus Protease

Background

TEV Protease is the 241 amino acid (aa), 27 kDa catalytic domain of the nuclear inclusion a (NIa) protein encoded by the potyvirus, tobacco etch virus (TEV). It may be used in biotechnology to cleave affinity tags from recombinant proteins, either co-translationally or in vitro following purification. Its high specificity and activity at a wide range of pH and ionic strength make TEV Protease more versatile than many other proteases used for the same purpose. Unlike factor Xa, enteropeptidase or thrombin, TEV Protease has not been found to cleave at unintended sites, even when present at a high concentration. TEV Protease is a 3C-type protease that cleaves substrates with a consensus sequence of ENLYFQG. Cleavage occurs between Q and G. Since the final aa remains on the cleaved protein where it could potentially affect structure or function, substitution of a variety of aa have been tested. In order of efficiency, S, A, M, Y, D, N, E, K or L may be effectively used in place of G. Several of the remaining aa may also vary, giving a final consensus sequence of ExxYF(M)Q(E)/G(S, A or others) where aa in parenthesis are alternatives and x is any aa. The autocatalytic site of NIa at S2256 has been mutated to an N for improved stability of the protease.

  1. Daros, J.-A. et al. (1999) J. Virol. 73:8732.
  2. Mondigler, M. and M. Ehrmann (1996) J. Bacteriol. 178:2986.
  3. Phan, J. et al. (2002) J. Biol. Chem. 277:50564.
  4. Kapust, R.B. et al. (2002) Biochem. Biophys. Res. Commun. 294:949.
  5. Kapust, R.B. et al. (2001) Protein Eng. 14:993.

Publications for TEV Protease (4469-TP)(1)

We have publications tested in 1 confirmed species: Rat.

We have publications tested in 1 application: Enzyme Assay.


Filter By Application
Enzyme Assay
(1)
All Applications
Filter By Species
Rat
(1)
All Species

Reviews for TEV Protease (4469-TP) (0)

There are no reviews for TEV Protease (4469-TP). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen

FAQs for TEV Protease (4469-TP) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Other Available Formats

Additional TEV Protease Products

Blogs on TEV Protease

There are no specific blogs for TEV Protease, but you can read our latest blog posts.
Recombinant Monoclonal Antibodies

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our Recombinant TEV Protease Protein, CF and receive a gift card or discount.

Bioinformatics

Uniprot