Product Class: Other

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

Product Introduction

α2-3,6,8 Neuraminidase catalyzes the hydrolysis of α2-3, α2-6, and α2-8 linked sialic acid residues from glycoproteins and oligosaccharides.

  • Recombinant enzyme with no detectable endoglycosidase or other exoglycosidases contaminating activities
  • Acts on both Neu5Ac and Neu5Gc
  • Fast digestion, removal of sialic acids in 5 minutes
Catalog # Size Concentration
P0720S 2000.0 units 50000 units/ml
P0720L 10000.0 units 50000 units/ml

Product Information

Description


 
Neuraminidase is the common name for Acetyl-neuraminyl hydrolase (Sialidase). This Neuraminidase catalyzes the hydrolysis of α2-3, α2-6, and α2-8 linked N-acetylneuraminic acid residues from glycoproteins and oligosaccharides.


Substrate Specificity:



Product Source

Cloned from Clostridium perfringens and overexpressed 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
  • P0720S     -20    
      α2-3,6,8 Neuraminidase P0720SVIAL -20 1 x 0.04 ml 50,000 units/ml
      GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
  • P0720L     -20    
      α2-3,6,8 Neuraminidase P0720LVIAL -20 1 x 0.2 ml 50,000 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-7-amino-4-methyl-coumarin (AMC), in 5 minutes 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
5 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Apparent: 43 kDa

Unit Assay Conditions

Two fold dilutions of α2-3,6,8 Neuraminidase 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 5 minutes. Separation of reaction products are visualized via thin layer chromatography (3).

References

  1. Roggentin, P. et al. (1988). FEBS Lett. 238(1), 31-34.
  2. Guan, C., New England Biolabs unpublished observations.
  3. 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 Neuraminidase (P0720)
  2. Endo-α-N-Acetylgalactosaminidase Application Note 1

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. Can I use α2-3,6,8 Neuraminidase in a double digest with Endo H(Hf) or PNGase F?
  2. Is α2-3,6,8 Neuraminidase active at higher pH levels?
  3. How much exoglycosidase should be used?
  4. Do detergents inhibit exoglycosidases/endoglycosidases?
  5. What is a good positive control for α2-3,6,8 Neuraminidase?
  6. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
  7. What are Glycosidases and their uses?
  8. Is it necessary to treat my glycoprotein concomitantly with Neuraminidase and O-Glycosidase?
  9. Do the NEB Neuraminidase enzymes cleave both N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) residues?
  10. How much exoglycosidase should be used?

Tech Tips

Don’t forget to include Neuraminidase in a digestion with O-Glycosidase. 

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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