Recombinant Influenza A Virus H5N1 Neuraminidase Protein, CF

R&D Systems | Catalog # 7597-NM

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems NS0-derived Recombinant Influenza A Virus H5N1 Neuraminidase Protein (7597-NM)
  • 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

NS0

Accession Number

Q6DPL2

Applications

Enzyme Activity
View all Viral Neuraminidase Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived influenza a virus h5n1 Viral Neuraminidase protein
Ser37-Lys449, with an N-terminal vasodilator-stimulated phosphoprotein tetramerization domain and a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Ser (tetramerization domain)

Predicted Molecular Mass

51 kDa

SDS-PAGE

58-65 kDa, reducing conditions

Activity

Measured by its ability to cleave a fluorogenic substrate, 2’-(4-Methylumbelliferyl)-alpha -D-N-acetylneuraminic acid.
The specific activity is >2,500 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation, and Storage

7597-NM
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: Viral Neuraminidase

Neuraminidase (NA) and hemagglutinin (HA) are the two predominant membrane glycoproteins found on the surface of an influenza virus particle. They are essential for the infectious cycle of the virus. HA recognizes and binds to the sialic acid on the host cell membrane to initiate a viral infection. NA cleaves the sialic acid at the end of the cycle, allowing the progeny virus to leave the host and initiate the next round of infection (1). In the early stage of an infection, NA may also assist in viral penetration of the mucus layer in the airway of a host. Nine subtypes of NA (N1 to N9) have been identified, all of which are believed to be tetrameric and share a basic structure consisting of a globular head, a thin stalk region, and a small hydrophobic region that anchors the protein in the virus membrane (2). Glycosylation has also found been important for the stability and activity of these enzymes (3, 4). According to a recent structure determination (5), there are two genetically distinct groups of neuraminidases from influenza type A viruses, with the N1 and N2 neuraminidases representing the two groups. Due to their critical role in the infectious cycle of a virus, influenza viral neuraminidases are frequently used as targets for drug design. In fact, both Tamiflu and Relenza, anti-influenza drugs, are neuraminidase inhibitors. Our recombinant H5N1 neuraminidase is based on the avian virus isolated from the 2004 outbreaks of the H5N1 virus in Vietnam and Thailand (6). H5N1 avian virus is one of the most lethal viruses in history (7, 8) with an accumulative death rate of 59% from 2003 to 2012 according to the World Health Organization (9). To produce active recombinant enzyme, a tetramerization domain from the vasodilator-stimulated phosphoprotein (10) was inserted at the N‑terminus to assist in oligomerization of the protein. We found that the activity of the recombinant H5N1 neuraminidase is activated by Ca2+ and inactivated by Zn2+, Cu2+ and Fe2+.

References

  1. Palese, P. & Compans, R. W. (1976) J. Gen. Virol. 33:159.
  2. Colman, P. M. et al. (1983) Nature 303:41.
  3. Wu, Z.L. et al. (2009) Biochem. Biophys. Res. Commun. 379:749.
  4. Sun, S. et al. (2012) PLoS one 7:e32119.
  5. Russell, R.J. et al. (2006) Nature 443:45.
  6. Govorkova, E.A. et al. (2005) J. Virol. 79:2191.
  7. Ducatez, M.F. et al. (2006) Nature 442:37.
  8. Peiris, J.S. et al. (2007) Clin. Microbiol. Rev. 20:243.
  9. http://www.who.int/influenza/human_animal_interface/EN_GIP_20120405NCumulativeNumberH5N1cases.pdf
  10. Kuhnel, K. et al. (2004) Proc. Natl. Acad. Sci. U. S. A. 101:17027.

Long Name

Neuraminidase

Alternate Names

NANH

UniProt

Q6DPL2

Additional Viral Neuraminidase Products

Product Documents for Recombinant Influenza A Virus H5N1 Neuraminidase 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.

Download SDS

Product Specific Notices for Recombinant Influenza A Virus H5N1 Neuraminidase Protein, CF

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

For research use only

Citations for Recombinant Influenza A Virus H5N1 Neuraminidase Protein, CF

Customer Reviews for Recombinant Influenza A Virus H5N1 Neuraminidase Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Influenza A Virus H5N1 Neuraminidase Protein, CF and earn rewards!

Have you used Recombinant Influenza A Virus H5N1 Neuraminidase 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
Amazon Gift Card

Protocols

View specific protocols for Recombinant Influenza A Virus H5N1 Neuraminidase Protein, CF (7597-NM):

Materials
  • Activation Buffer: 50 mM MES, 500 mM NaCl, 30 mM CaCl2, pH 6.5
  • Assay Buffer: 50 mM MES, 500 mM NaCl, 5 mM CaCl2, pH 6.5
  • Recombinant Influenza A Virus H5N1 Neuraminidase (rvH5N1 Neuraminidase) (Catalog # 7597-NM)
  • Substrate: 4-Methylumbelliferyl-alpha -D-N-acetylneuraminic acid (Sigma, Catalog # M8639), 10 mM stock  in DMSO
  • F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
  • Fluorescent Plate Reader (Model: Gemini EM by Molecular Devices) or equivalent
  1. Dilute rvH5N1 Neuraminidase to 100 μg/mL in Activation Buffer.
  2. Incubate for 24 hours at 37 °C to activate.
  3. Dilute activated rvH5N1 Neuraminidase to 0.5 ng/μL in Assay Buffer.
  4. Dilute Substrate to 400 µM in Assay Buffer.
  5. Load into a black well plate 50 µL of the 0.5 ng/μL rvH5N1 Neuraminidase and start the reaction by adding 50 µL of 400 µM Substrate. Include a Substrate Blank containing Assay Buffer in place of rvH5N1 Neuraminidase.
  6. Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively, in kinetic mode for 5 minutes.
  7. 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 4-Methylumbelliferone (Sigma, Catalog # M1381).

Per Well:
  • rvH5N1 Neuraminidase: 0.025 µg
  • Substrate: 200 µM

FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes