Recombinant M. viridifaciens Neuraminidase Protein, CF

R&D Systems | Catalog # 5084-NM

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Key Product Details

  • R&D Systems E. coli-derived Recombinant M. viridifaciens Neuraminidase Protein (5084-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

E. coli

Accession Number

BAA00852

Applications

Enzyme Activity
View all Bacterial Neuraminidase Proteins and Enzymes »

Product Specifications

Source

E. coli-derived m. viridifaciens Bacterial Neuraminidase protein
Pro42-Gly403, with an N-terminal Met and 6-His tag
Accession # BAA00852

Purity

>95%, 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

Met

Predicted Molecular Mass

40 kDa

SDS-PAGE

42 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 >25,000 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation, and Storage

5084-NM
Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
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: Bacterial Neuraminidase

Neuraminidase is the common name for N-acetyl-neuraminyl hydrolase (Sialidase). Micromonospora viridifaciens Neuraminidase catalyzes the hydrolysis of alpha 2-3 and alpha 2-6 and alpha 2-8 linked N-acetyl-neuraminic acid residues from glycoproteins and glycolipids (1). The architecture of the full-length protein includes a canonical neuraminidase enzymatic domain, a linker domain and a C-terminal galactose binding domain (2). The full-length protein contains 647 amino acids and has a molecular weight of 68 kDa. It is easily degraded to 52 kDa and 41 kDa fragments (3). The expressed enzyme only contains the enzymatic domain.

References

  1. Aisaka, K. & Uwajima, T. (1987) FEBS Microbiol. Lett. 44:289.
  2. Gaskell, A. et al. (1995) Structure 3:1197.
  3. Sakurada, K. et al. (1992) J. Bacteriol. 174:6896.

UniProt

BAA00852

Additional Bacterial Neuraminidase Products

Product Documents for Recombinant M. viridifaciens Neuraminidase Protein, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant M. viridifaciens Neuraminidase Protein, CF

For research use only

Related Research Areas

Glycobiology-related Enzymes

Citations for Recombinant M. viridifaciens Neuraminidase Protein, CF

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Protocols

View specific protocols for Recombinant M. viridifaciens Neuraminidase Protein, CF (5084-NM):

Materials
  • Assay Buffer: 50 mM Sodium Acetate, 150 mM NaCl, pH 4.5
  • Recombinant M. viridifaciens Neuraminidase (rMvNA) (Catalog # 5084-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 rMvNA to 0.2 ng/µL in Assay Buffer.
  2. Dilute Substrate to 400 µM with Assay Buffer.
  3. Load into plate 50 µL of 0.2 ng/µL rMvNA and start the reaction by adding 50 µL of 400 µM Substrate. Include a Substrate Blank containing 50 µL of Substrate and 50 µL of Assay Buffer.
  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 (pmoles/min/µg) =

 Adjusted Vmax* (RFU/min) x Conversion Factor** (pmole/RFU)
amount of enzyme (µg)

     *Adjusted for Substrate Blank

     **Derived using calibration standard 4-Methylumbelliferone (Sigma, Catalog # M1381).

  • rMvNA: 0.010 µg
  • Substrate: 200 µM

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