Recombinant C. perfringens Neuraminidase Protein, CF

• Reactivity: Cp
• Applications: Enzyme Activity
• Format: Carrier-Free

Recombinant C. perfringens Neuraminidase Protein, CF Summary

• Details of Functionality: Measured by its ability to cleave a fluorogenic substrate, 2’-(4-Methylumbelliferyl)-alpha -D-N-acetylneuraminic acid. The specific activity is >80,000 pmol/min/µg, as measured under the described conditions.
• Source: E. coli-derived c. perfringens Bacterial Neuraminidase protein
  Cys2-Gln382, with an N-terminal Met and 6-His tag
• Accession #: P10481
• N-terminal Sequence: Met
• Protein/Peptide Type: Recombinant Enzymes
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <1.0 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Enzyme Activity
• Theoretical MW: 44 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 40 kDa, reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Supplied as a 0.2 μm filtered solution in Tris, NaCl and EDTA.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Assay Procedure:
  • Assay Buffer: 50 mM MES, 100 mM NaCl, 0.05% (w/v) Brij-35, pH 6.5
  • Recombinant C. perfringens Neuraminidase (rCpNA) (Catalog # 5080-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 rCpNA to 0.04 ng/µL in Assay Buffer.
  2. Dilute Substrate to 400 µM in Assay Buffer.
  3. Load 50 µL of 0.04 ng/µL rCpNA into a plate, and start the reaction by adding 50 µL of 400 µM 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 (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).

  Per Well:
  • rCpNA: 0.002 µg
  • Substrate: 200 µM

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant C. perfringens Neuraminidase Protein, CF

• Bacterial Neuraminidase

Background

Neuraminidase and Sialidase are the common names for N‑acetyl‑neuraminyl hydrolase (1-3). The enzyme catalyzes the hydrolysis of alpha 2-3 and alpha 2-6 linked N‑acetyl‑neuraminic acid residues from glycoproteins and glycolipids. It has little activity against the alpha 2-8 linked N‑acetyl‑neuraminic acid residues.

1. Christensen, S. et al. (2005) Biotechnol. Appl. Biochem. 41:225.
2. Roggentin, P. et al. (1988) FEBS Lett. 238:31.
3. Corfield, A. et al. (1983) Biochim. Biophys. Acta 744:121.

Publications for Bacterial Neuraminidase (5080-NM)(1)

Publication using 5080-NM

• ZL Wu, JM Ertelt Fluorescent glycan fingerprinting of SARS2 spike proteins Scientific Reports, 2021;11(1):20428. 2021 [PMID: 34650101] (Bioassay, Human)

Bioinformatics

• Uniprot:
  • P10481(C. perfringens)