Infection of mosquito-borne West Nile Virus can cause severe neurological disease and can be epidemic. Two non-structural proteins, NS3 and NS2b, play an essential role in viral replication and are therefore a potential target for treatment and prevention of West Nile Virus disease. NS3 consists of a trypsin-like serine protease with a catalytic triad (His51, Asp75, Ser135) and a putative helicase. Requiring NS2b as the co‑factor, NS3 protease processes viral polyprotein precursor (1, 2). The purified recombinant protein consists of three forms: the full-length fusion protein, the N-terminal NS2b, and the C-terminal NS3 with the G4SG4 linker. NS3 protease has a relatively narrow substrate specificity that prefers Arg in P1 and Lys in P2. The purified recombinant protein has autocatalytic activity that can lead to protein degradation. It is therefore important to store the sample below -20 °C and to keep on ice while working with the sample.
Recombinant West Nile Virus NS3 Protease Protein, CF
R&D Systems | Catalog # 2907-SE
Key Product Details
- R&D Systems E. coli-derived Recombinant West Nile Virus NS3 Protease Protein (2907-SE)
- 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
Accession Number
Applications
Product Specifications
Source
| MHHHHHH | NS2b (Ser1423-Lys1470) (Ser1435Thr) Accession # YP_001527877.1 |
GGGGSGGGG | West Nile Virus NS3 |
|
| GGGGSGGGG | West Nile Virus NS3 (Gly1506-Leu1689) Accession # YP_001527877.1 |
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| N-terminus | C-terminus | |||
Purity
Endotoxin Level
N-terminal Sequence Analysis
Predicted Molecular Mass
SDS-PAGE
Activity
The specific activity is >750 pmol/min/µg, as measured under the described conditions.
Formulation, Preparation, and Storage
2907-SE
| Formulation | Lyophilized from a 0.2 μm filtered solution in Tris and NaCl. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile, deionized water.
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| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Calculators
Background: wnvNS3 Protease
References
- Nall, T.A. et al. (2004) J. Biol. Chem. 279:48535.
- Chappell, K.J. et al. (2005) J. Biol. Chem. 274:2896.
Long Name
Entrez Gene IDs
Gene Symbol
UniProt
Additional wnvNS3 Protease Products
Product Documents for Recombinant West Nile Virus NS3 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 West Nile Virus NS3 Protease Protein, CF
For research use only
Related Research Areas
Citations for Recombinant West Nile Virus NS3 Protease Protein, CF
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Protocols
View specific protocols for Recombinant West Nile Virus NS3 Protease Protein, CF (2907-SE):
- Assay Buffer: 50 mM Tris, 30% (v/v) Glycerol, pH 9.5
- Recombinant Viral wnvNS3 Protease (Catalog # 2907-SE)
- Substrate: L-PYROGlu-Arg-Thr-Lys-Arg-AMC (pERTKR-AMC) (Catalog # ES013)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Dilute rwnvNS3 Protease to 1 ng/µL in Assay Buffer.
- Dilute Substrate to 40 µM in Assay Buffer.
- In a plate load 50 µL of 1 ng/µL rwnvNS3 Protease, and start the reaction by adding 50 µL of 40 µM Substrate to wells. Include a Substrate Blank containing 50 µL Assay Buffer and 50 µL of 40 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Calculate specific activity:
|
Specific Activity (pmol/min/µg) = |
Adjusted Vmax* (RFU/min) x Conversion Factor** (pmol/RFU) |
| amount of enzyme (µg) |
*Adjusted for Substrate Blank
**Derived using calibration standard 7-Amino, 4-Methyl Coumarin (AMC) (Sigma, Catalog # A-9891).
Per Well:
- rwnvNS3 Protease: 0.05 µg
- Substrate: 20 µM