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Exoglycosidases >
α2-3 Neuraminidase |
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Substrate Specificity:
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).
Source:
Cloned from Salmonella typhimurium LT2 and overexpressed in E. coli (1).
Reagents Supplied:
G4 Reaction Buffer (10X)
BSA (100X)
Enzyme Properties
Molecular Weight:
Apparent: 41 kDa
Specific Activity:
11,300,000 units/mg
Reaction & Storage Conditions
Reaction Conditions:
1X G4 Reaction Buffer
Supplemented with 100 μg/ml Bovine Serum Albumin
Incubate at
37°C.
1X G4 Reaction Buffer:
50 mM sodium citrate
100 mM NaCl
pH 6.0 @ 25°C
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.
Two fold dilutions of α2-3 Neuraminidase are incubated with 1 nmol AMC-labeled substrate and 1X G4 Buffer, 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).
Concentration:
50,000 units/ml
Storage Conditions:
20 mM Tris-HCl
50 mM NaCl
5 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
4°C
Avoid repeated freeze/thaw cycles.
FAQs
- Can I use α2-3 Neuraminidase in a cocktail with Endo H(Hf) or PNGase F?
- What is the difference between Neuraminidase and α2-3 Neuraminidase?
- Do detergents inhibit exoglycosidases/endoglycosidases?
- How much exoglycosidase should be used?
- 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 excontaminating exoglycosidase or proteolytic activity could be detected.
Protease Activity:
After incubation of 500 units of α2-3 Neuraminidase with a standard mixture of proteins
for 2 hours at 30ºC, no proteolytic activity could be detected by SDS-PAGE.
Glycosidase Assay:
500 units of α2-3 Neuraminidase were incubated with 0.1 mM of flourescently-labeled oligosaccharides and glycopeptides, in a 10 µl reaction for 20 hours at 37°C. The reaction products were analyzed by TLC for digestion of substrate.
No other glycosidase activities were detected (ND) with the following substrates:
β-N-Acetyl-glucosaminidase: GlcNAcβ1-4GlcNAcβ1-4GlcNAc-AMC ND
α-Fucosidase: Fucα1-2Galβ1-4Glc-AMCGalβ1-4 (Fucα1-3)GlcNAcβ1-3Galβ1-4Glc-AMC ND
β-Galactosidase: Galβ1-3GlcNAcβ1-4Galβ1-4Glc-AMC ND
α-Galactosidase: Galα1-3Galβ1-4Galα1-3Gal-AMC ND
α-Mannosidase: Manα1-3Manβ1-4GlcNAc-AMC Manα1-6Manα1-6(Manα1-3)Man-AMC ND
β-Glucosidase: Glcβ1-4Glcβ1-4Glc-AMC ND
β-Xylosidase: Xylβ1-4Xylβ1-4Xylβ1-4Xyl-AMC ND
β-Mannosidase: Manβ1-4Manβ1-4Man-AMC ND
Endo F1, F2, H: Dansylated invertase high mannose. ND
Endo F2, F3: Dansylated fibrinogen biantennary. ND
PNGase F: Fluoresceinated fetuin triantennary. ND
References
- Hoyer, et al. (1991) J. Biochem., 110, 462-467.
- Monks, B., New England Biolabs, unpublished observations.
- Wong-Madden, S.T. and Landry, D. (1995) Glycobiology, 5, 19-28.
Reagents Sold Separately
BSA
Companion Products
Exoglycosidase Reaction Buffer Pack
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