Recombinant Influenza A Virus H1N1 Neuraminidase Protein, CF

R&D Systems | Catalog # 4858-NM

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Key Product Details

  • R&D Systems Sf 21 (baculovirus)-derived Recombinant Influenza A Virus H1N1 Neuraminidase Protein (4858-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

Sf 21 (baculovirus)

Accession Number

AAF77036

Applications

Enzyme Activity
View all Viral Neuraminidase Proteins and Enzymes »

Product Specifications

Source

Spodoptera frugiperda, Sf 21 (baculovirus)-derived influenza a virus h1n1 Viral Neuraminidase protein
Ser37-Lys469, with an N-terminal 6-His tag

Purity

>80%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His

Predicted Molecular Mass

48 kDa

SDS-PAGE

57 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

4858-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 major membrane glycoproteins found on the surface of influenza virus. HA is a lectin that binds sialic acid on host cell membrane. NA is a sialic acid hydrolase that specifically clips off terminally located sialic acid on host cell surface. The two proteins are essential for the infectious cycle of the influenza virus. During initial infection, an influenza virus will hold onto an epithelial cell through HA-sialic acid interaction. At the end of an infectious cycle, the NA will cleave the sialic acid on the host cell membrane, releasing the formed viral particle from the HA-sialic acid bondage (1). The neuraminidase activity is also thought to help the virus penetrate mucus. Nine subtypes of NA have been identified, all of which are tetrameric and share a common structure consisting of a globular head, a thin stalk region, and a small hydrophobic region that anchors the protein in the virus membrane (2). The purified rvH1N1NA consists of amino acid residues 37 to 469 as deduced from the 1918 Spanish flu virus NA (A/Bervig_Mission/1/18) (3). It has a distinct N-glycan profile and is resistant to trypsin digestion (4).

References

  1. Palese, P. & Compans, R. W. (1976) J. Gen. Virol. 33:159.
  2. Colman, P. M. et al. (1983) Nature 303:41.
  3. Reid, A. (2000) Proc. Natl. Acad. Sci. USA 97:6785.
  4. Wu, Z.L. et al. (2009) Biochem. Biophys. Res. Comm. 379:749.

Long Name

Neuraminidase

Alternate Names

NANH

UniProt

AAF77036 (Viral)

Additional Viral Neuraminidase Products

Product Documents for Recombinant Influenza A Virus H1N1 Neuraminidase Protein, CF

Certificate of Analysis

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Product Specific Notices for Recombinant Influenza A Virus H1N1 Neuraminidase Protein, CF

For research use only

Citations for Recombinant Influenza A Virus H1N1 Neuraminidase Protein, CF

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Protocols

View specific protocols for Recombinant Influenza A Virus H1N1 Neuraminidase Protein, CF (4858-NM):

Materials
  • Assay Buffer: 50 mM Tris, 5 mM CaCl2, 200 mM NaCl, pH 7.5
  • Recombinant Influenza A Virus H1N1 Neuraminidase (rH1N1 Neuraminidase) (Catalog # 4858-NM)
  • Substrate: 2’-(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: SpectraMax Gemini EM by Molecular Devices) or equivalent
  1. Dilute rH1N1 Neuraminidase to 0.05 ng/µL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer.
  3. Load 50 µL of 0.05 ng/µL rH1N1 Neuraminidase into the plate, and start the reaction by adding 50 µL of 400 µM Substrate. As a Substrate Blank load 50 µL of Assay Buffer and 50 µL of Substrate.
  4. Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively in kinetic mode for 5 minutes.
  5. 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:
  • rH1N1 Neuraminidase: 0.0025 µg
  • Substrate: 200 µM

FAQs

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