• My NEB
  • Print
  • PDF
  • Neuraminidase

    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-acetyl-neuraminic acid residues from glycoproteins and oligosaccharides.


    Substrate Specificity:




    Product Source

    Cloned from Clostridium perfringens and overexpressed in E. coli

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    G1 Reaction Buffer10X

    Properties and 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.

    Two fold dilutions of Neuraminidase are incubated with 1 nmol AMC-labeled substrate and 1X G1 Buffer 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).

    Reaction Conditions

    1X G1 Reaction Buffer
    Incubate at 37°C

    1X G1 Reaction Buffer:
    50 mM sodium citrate
    pH 6 @ 25°C

    Storage Temperature

    -20°C

    Storage Conditions

    20 mM Tris-HCl
    50 mM NaCl
    5 mM Na2EDTA
    pH 7.5 @ 25°C

    Heat Inactivation

    65°C for 10 min

    Molecular Weight

    Apparent: 43 kDa

    Quality Control

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Glycosidase Activity (TLC):
      The product is tested in multiple reactions, each containing a fluorescently-labeled oligosaccharide or glycopeptide.  The reaction products are analyzed by TLC for digestion of substrate. No contaminating exoglycosidase or endoglycosidase activity is detected.
    • Protease Activity (SDS-PAGE):

      The product is tested for protease activity by incubation with a standard mixture of proteins resulting in no detectable degradation of the proteins as determined by SDS-PAGE.

    • Protein Purity (SDS-PAGE):
      The physical purity is assessed by comparing contaminating protein bands in a concentrated sample to the protein of interest band in a sample of known dilution. The purity is determined by SDS-PAGE.

    Notes

    1. This enzyme shows a preference for α2-3 and α2-6 linkages over α2-8 linkages.

    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.

    Supporting Documents

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.

    Datacards

    The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.
    1. Is it necessary to treat my glycoprotein concomitantly with Neuraminidase and O-Glycosidase?
    2. Can I use Neuraminidase in a double digest with Endo H(Hf) or PNGase F?
    3. Is Neuraminidase active at higher pH levels?
    4. How much exoglycosidase should be used?
    5. Do detergents inhibit exoglycosidases/endoglycosidases?
    6. What is a good positive control for neuraminidase?
    7. What are Glycosidases and their uses?
    1. Typical Reaction Conditions for Neuraminidase (P0720)
    2. Endo-α-N-Acetylgalactosaminidase Application Note 1

    Usage Guidelines & Tips

    Application Notes

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