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α2-3,6,8,9 Neuraminidase A

Catalog # Concentration Size List Price Quantity Your Price
P0722L 20000 units/ml 4000 units $465.00
$418.50
P0722S 20000 units/ml 800 units $117.00
$105.30
Catalog # Size List Price Your Price
P0722L 4000 units $465.00
$418.50
P0722S 800 units $117.00
$105.30
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

α2-3,6,8,9 Neuraminidase A is a broad specificity sialidase, which cleaves linear and branched non-reducing terminal sialic acid residues from glycoproteins, glycopeptides, and oligosaccharides. It can be used for glycan analysis and characterization and intact glycoprotein remodeling, in vitro and in vivo.

  • Recombinant enzyme with no detectable endoglycosidase or other exoglycosidases contaminating activities
  • Acts on both Neu5Ac and Neu5Gc
  • Double digest with other exoglycosidases and endoglycosidases
  • Tolerant of moderate levels (0.5-1.0%) of detergents
  • ≥95% purity, as determined by SDS-PAGE and intact ESI-MS
  • Optimal activity and stability for up to 24 months


 
Neuraminidase is the common name for Acetyl-neuraminyl hydrolase (Sialidase). α2-3,6,8,9 Neuraminidase A catalyzes the hydrolysis of all linear and branched non-reducing terminal sialic acid residues from glycoproteins and oligosaccharides. The enzyme releases α2-3 and α2-6 linkages at a slightly higher rate than α2-8 and α2-9 linkages.

Specificity


Detailed Specificity
α2-3,6,8,9 Neuraminidase A will cleave branched sialic acid residues that are linked to an internal residue. This oligosaccharide from fetuin is an example of a side-branch sialic acid residue that can efficiently be cleaved (1).


Product Source
Cloned from Arthrobacter ureafaciens and expressed in E. coli.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  α2-3,6,8,9 Neuraminidase A P0722SVIAL -20 1 x 0.04 ml 20,000 units/ml
  GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
  α2-3,6,8,9 Neuraminidase A P0722LVIAL -20 1 x 0.2 ml 20,000 units/ml
  GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of the terminal α-Neu5Ac from 1 nmol Neu5Acα2-3Galβ1- 3GlcNAcβ1-3Galβ1-4Glc-AMC, in 1 hour at 37°C in a total reaction volume of 10 μl.

Reaction Conditions

1X GlycoBuffer 1
Incubate at 37°C

1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)

Storage Buffer

50 mM NaCl
20 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Apparent: 100000 daltons

Specific Activity

316,000 units/mg

Unit Assay Conditions

Two fold dilutions of α2- 3,6,8,9 Neuraminidase A are incubated with 1 nmol AMC-labeled substrate and 1X GlycoBuffer 1 in a 10 μl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via thin layer chromatography (2).

Companion Products
Notes
  • Reactions may be scaled-up linearly to accommodate larger reaction volumes.
  • The amount of exoglycosidase enzyme required varies when different substrates are used. Start with 1–2 μl for 1 μg of glycoprotein or 100 nM of oligosaccharide for one hour in a 10–25 μl reaction. If there is still undigested material, let the reaction go overnight.
  • Higher concentrations of enzyme as well as longer incubation times may be necessary for cleavage of branched structures.
References
  • Iwamori, M., Ohta, Y., Uchida, Y. and Tsukada, Y. (1997). Glycocon. J. 1, 67-73.
  • Wong-Madden, S. T. and Landry, D. (1995). Glycobiology. 5, 19-28.
Web Tools
Citations
Publications
  • Albrecht, S., Vainauskas, S., Stöckmann, H., McManus, C., Taron, C.H. and Rudd, P.M. (2016). Comprehensive Profiling of Glycosphingolipid Glycans Using a Novel Broad Specificity Endoglycoceramidase in a High-Throughput Workflow. Anal Chem. May 3;88(9), 4795-802. Anal Chem..PubMedID: 27033327
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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