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.
Recombinant TEV Protease Protein, CF
R&D Systems | Catalog # 4469-TP
Loading...
Key Product Details
- R&D Systems E. coli-derived Recombinant TEV Protease Protein (4469-TP)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Source
E. coli
Accession Number
Applications
Enzyme Activity
Loading...
Product Specifications
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 | ||
Purity
>80%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Ala
Predicted Molecular Mass
28.5 kDa
SDS-PAGE
25-30 kDa, reducing conditions
Activity
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.
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.
Reviewed Applications
Read 1 review rated 3 using 4469-TP in the following applications:
Formulation, Preparation, and Storage
4469-TP
| Formulation | Supplied as a 0.2 μm filtered solution in Tris, NaCl, EDTA, DTT and Glycerol. |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: TEV Protease
References
- Daros, J.-A. et al. (1999) J. Virol. 73:8732.
- Mondigler, M. and M. Ehrmann (1996) J. Bacteriol. 178:2986.
- Phan, J. et al. (2002) J. Biol. Chem. 277:50564.
- Kapust, R.B. et al. (2002) Biochem. Biophys. Res. Commun. 294:949.
- Kapust, R.B. et al. (2001) Protein Eng. 14:993.
Long Name
Tobacco Etch Virus Protease
Alternate Names
NIa
UniProt
Additional TEV Protease Products
Product Documents for Recombinant TEV Protease Protein, CF
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Recombinant TEV Protease Protein, CF
For research use only
Related Research Areas
Citations for Recombinant TEV Protease Protein, CF
Powered by Bioz
Customer Reviews for Recombinant TEV Protease Protein, CF (1)
3 out of 5
1 Customer Rating
Have you used Recombinant TEV Protease Protein, CF?
Submit a review and receive an Amazon gift card!
$25/€18/£15/$25CAN/¥2500 Yen for a review with an image
$10/€7/£6/$10CAN/¥1110 Yen for a review without an image
Submit a review
Showing
1
-
1 of
1 review
Showing All
Filter By:
-
Application: Protein IsolationVerified Customer | Posted 10/09/2020
There are no reviews that match your criteria.
Protocols
View specific protocols for Recombinant TEV Protease Protein, CF (4469-TP):
Materials
- 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
- Dilute rvTEV Protease to 0.02 mg/mL in Assay Buffer.
- Dilute Substrate to 0.5 mg/mL in Assay Buffer.
- 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.
- Incubate all three mixtures (temperature equilibration), as well as the diluted rvTEV Protease, for 15 minutes at room temperature.
- Add 10 µL diluted rvTEV Protease to the reaction mixture, excluding the controls, for a final volume of 50 µL in each reaction.
- 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.
- 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.
- Analyze the cleavage by SDS-PAGE followed by protein staining.
- rvTEV Protease: 0.2 µg
- Substrate: 10 µg