α2-3 Neuraminidase

This product has been discontinued and replaced by P0743S/L .
  • This product was discontinued on 01/01/2017
recombinant unique buffer incubation temp heat inactivation bsa
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Discontinued Products

Description

α2-3 Neuraminidase is a highly specific exoglycosidase that catalyzes the hydrolysis of α2-3 and, at a much lower rate, α2-6 linked N-acetyl-neuraminic acid residues from oligosaccharides. This enzyme has a 260-fold preference for α2-3 sialyl linkages over α2-6 sialyl linkages and shows only trace activity against α2-8 sialyl linkages (1).

Substrate Specificity:

Product Source

Cloned from Salmonella typhimurium LT2 and overexpressed in E. coli (1).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
GlycoBuffer 1-2010X
Purified BSA-2010X

Properties and Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of the αNeu5Ac from 1 nmol of Neu5Acα2-3Galβ1-3GlcNAcβ1-3Galβ1-4Glc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 µl.

Unit Definition Assay
Two fold dilutions of α2-3 Neuraminidase are incubated with 1 nmol AMC-labeled substrate and 1X GlycoBuffer 1, supplemented with 100 µg/ml BSA, in a 10 µl reaction. The reaction mix is incubated for 1 hour at 37°C. Separation of reaction products are visualized via thin layer chromatography (3).

Reaction Conditions

1X GlycoBuffer 1
Supplement with 100 μg/ml Purified BSA
Incubate at 37°C

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

Storage Temperature

4°C

Storage Conditions

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

Heat Inactivation

65°C for 10 min

Molecular Weight

Apparent: 41 kDa

References

  1. Hoyer, et al. (1991). J. Biochem. 110, 462-467.
  2. Monks, B., New England Biolabs Inc. unpublished observation.
  3. Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.

FAQs

  1. Can I use α2-3 Neuraminidase in a cocktail with Endo H(Hf) or PNGase F?
  2. What is the difference between Neuraminidase and α2-3 Neuraminidase?
  3. How much exoglycosidase should be used?
  4. 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?
  5. What are glycosidases and their uses?

Tech Tips

α2-3Neuraminidase can be used in double digests with PNGase F, Endo H and O-Glycosidase (under native or denaturing conditions).

Neuraminidase shows normal activity in the presence of 1% BME, and 0.5% SDS.

Repeated freeze-thaw cycles may reduce activity. Recommended storage temperature is 4°C.

Using 1-2 µl is a good starting point for a 1 hr incubation of 1µg of glycoprotein or 100 nM of oligosaccharide.

Protocols

  1. Typical Reaction Conditions for α2-3 Neuraminidase (P0728)

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.

Feature Articles

Usage Guidelines & Tips

Interactive Tools

Quality Control

Quality Assurance Statement

  • No contaminating exoglycosidase or proteolytic activity could be detected.

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. Further information regarding NEB product quality can be found here.
  • 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.

Certificate of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

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]

Safety Data Sheet

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

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.