Recombinant M. viridifaciens Neuraminidase Protein, CF

Catalog #: 5084-NM Datasheet / COA / SDS

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5084-NM-010

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Recombinant M. viridifaciens 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 >25,000 pmol/min/µg, as measured under the described conditions.

Source

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

Accession #

BAA00852

N-terminal Sequence
Analysis

Met

Predicted Molecular Mass

40 kDa

SDS-PAGE

42 kDa, reducing conditions

Product Datasheets

5084-NM

Product Datasheet

COA

SDS

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.

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.

Assay Procedure

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

Dilute rMvNA to 0.2 ng/µL in Assay Buffer.
Dilute Substrate to 400 µM with Assay Buffer.
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.
Read at excitation and emission wavelengths of 365 nm and 445 nm (top read), respectively, in kinetic mode for 5 minutes.
Calculate specific activity:

Specific Activity (pmoles/min/µg) =	Adjusted V_max* (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

Reconstitution Calculator

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