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Exoglycosidases >
Neuraminidase |
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Substrate Specificity:
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.
Source:
Cloned from Clostridium perfringens and overexpressed in E. coli
Reagents Supplied:
G1 Reaction Buffer (10X)
Enzyme Properties
Molecular Weight:
Apparent: 43 kDa
Specific Activity:
140,000 units/mg
Reaction & Storage Conditions
Reaction Conditions:
1X G1 Reaction Buffer
Incubate at
37°C.
1X G1 Reaction Buffer:
50 mM sodium citrate
pH 6.0 @ 25°C
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).
Concentration:
50,000 units/ml
Storage Temperature:
-20°C
Notes
Usage notes:- This enzyme shows a preference for α2-3 and α2-6 linkages over α2-8 linkages.
FAQs
- Can I use Neuraminidase in a double digest with Endo H(Hf) or PNGase F?
- Is Neuraminidase active at higher pH levels?
- What is the difference between Neuraminidase and α2-3 Neuraminidase?
- How much exoglycosidase should be used?
- Do detergents inhibit exoglycosidases/endoglycosidases?
- What is a good positive control for neuraminidase?
- Do detergents inhibit exoglycosidases/endoglycosidases?
- What are Glycosidases and their uses?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
No contaminating endoglycosidase, exoglycosidase, or proteolytic activity could be detected.
Protease Activity:
After incubation of 500 units of Neuraminidase with a standard mixture of proteins
for 2 hours at 30ºC, no proteolytic activity could be detected by SDS-PAGE.
Exo/endoglycosidase Activity:
After incubation of 500 units of Neuraminidase
with a variety of p-nitrophenyl-glycoside substrate and fluorescently-labeled oligosaccharides for 20 hours at 37ºC, no
contaminating exoglycosidase or endoglycosidase activities could be detected by spectrophotometric analysis or thin layer chromatography.
Contaminating activities tested for include ()
References
- Roggentin, P. et al. (1988) FEBS Lett., 238(1), 31-34.
- Guan, C., New England Biolabs, unpublished observations.
- Wong-Madden, S.T. and Landry, D. (1995) Glycobiology, 5, 19-28.
Companion Products
Exoglycosidase Reaction Buffer Pack
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