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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).
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)
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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 [email protected] or fill out the Technical Support Form for appropriate document.)
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