Product Class: Other

α2-3,6,8,9 Neuraminidase A
NEBU cloned at NEB recombinant 37 65 Heat

Product Introduction

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
Catalog # Size Concentration
P0722S 800.0 units 20000 units/ml
P0722L 4000.0 units 20000 units/ml

Product Information

Description


 
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.
This product is related to the following categories:
Exoglycosidases Products,
Proteome Analysis Products,
This product can be used in the following applications:
Expression Systems,
Glycan Sequencing,
Proteomics,
Recombinant Glycoprotein Expression,
Glycoprotein Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • P0722S     -20    
      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
  • P0722L     -20    
      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).

Product Notes

  1. Reactions may be scaled-up linearly to accommodate larger reaction volumes.
  2. 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.
  3. Higher concentrations of enzyme as well as longer incubation times may be necessary for cleavage of branched structures.

References

  1. Iwamori, M., Ohta, Y., Uchida, Y. and Tsukada, Y. (1997). Glycocon. J. 1, 67-73.
  2. Wong-Madden, S. T. and Landry, D. (1995). Glycobiology. 5, 19-28.

Protocols, Manuals & Usage

Protocols

  1. Typical Reaction Conditions for α2-3,6,8,9 Neuraminidase A (P0722)

Application Notes

Tools & Resources

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the difference between the four Neuraminidase enzymes sold by NEB: α2-3,6,8 Neuraminidase, α2-3,6,8,9 Neuraminidase A, α2-3 Neuraminidase and α2-3 Neuraminidase S?
  2. What is a good positive control for α2-3,6,8,9 Neuraminidase A?
  3. Can I use α2-3,6,8,9 Neuraminidase A in a double digest with other exoglycosidases and/or endoglycosidases?
  4. Can I use α2-3,6,8,9 Neuraminidase A in a double digest with Endo H/Hf, O-Glycosidase or PNGase F?
  5. Does this enzyme require denaturing conditions to act on glycoproteins?
  6. What is a good method to re-purify a glycan or glycopeptide after exoglycosidase treatment?
  7. Do detergents inhibit glycosidases?
  8. What are Glycosidases and their uses?
  9. Do the NEB Neuraminidase enzymes cleave both N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) residues?
  10. Is it necessary to treat my glycoprotein concomitantly with Neuraminidase and O-Glycosidase?
  11. How much exoglycosidase should be used?
  12. How much exoglycosidase should be used?

Tech Tips

  • Don’t forget to include Neuraminidase in a digestion with O-Glycosidase. 
  • f you are trying to cleave an oligosaccharide with a branched sialic acid residue you will want to use α2-3,6,8,9 Neuraminidase A

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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