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Recombinant C. perfringens Neuraminidase Protein, CF

Catalog #: 5080-NM
1 Citation Datasheet
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5080-NM-010
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Recombinant C. perfringens Neuraminidase Protein, CF Summary

Learn more about Fluorescent Glycan Labeling and Detection

Product Specifications

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.
Activity
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
Analysis
Met
Predicted Molecular Mass
44 kDa
SDS-PAGE
40 kDa, reducing conditions

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5080-NM

Product Datasheet

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

5080-NM

Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and EDTA.
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.

Assay Procedure

Materials
  • 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

Background: Bacterial Neuraminidase

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.

References
  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.
Alternate Names
Bacterial Neuraminidase

Citation for Recombinant C. perfringens Neuraminidase Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Fluorescent glycan fingerprinting of SARS2 spike proteins
    Authors: ZL Wu, JM Ertelt
    Scientific Reports, 2021-10-14;11(1):20428.
    Species: Human
    Sample Types: Recombinant Protein
    Applications: Bioassay

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