NEB, E0540S, O-Glycosidase & α2-3,6,8 Neuraminidase Bundle
This bundle contains 1 vial each of O-Glycosidase (
Catalog Number: E0540S
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Category
Pack Size1 vial
TagsGlycobiology
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| <b>Unit Definition</b> One unit of O-Glycosidase is defined as the amount of enzyme required to remove 0.68 nmol of O-linked disaccharide from 5 mg of Neuraminidase digested, non-denatured fetuin in 1 hour at 37°C in a total reaction volume of 100 μl (1 unit of both O-Glycosidase and PNGase F will remove equivalent molar amounts of O-linked disaccharides and N-linked oligosaccharides, respectively). Non-denaturing Unit Definition of O-Glycosidase | Two fold serial dilutions of O-Glycosidase are added to a reaction mixture of 5 mg of Neuraminidase digested fetuin with 1X GlycoBuffer 2. The reaction is then incubated at 37°C for 1 hour. O-linked disaccharide carbohydrates are determined by Morgan and Elson Assay (4). Note: Under denaturing conditions the enzyme activity is increased two-fold. This observation is substrate dependent. Unit Definition of Neuraminidase: One unit of Neuraminidase 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. 1X Glycoprotein Denaturing Buffer 0.5% SDS 40 mM DTT 1X NP-40 1% NP-40 in MilliQ-H 2 O <b>Reaction Conditions</b> 1X GlycoBuffer 2 Incubate at 37°C 1X GlycoBuffer 2 50 mM Sodium Phosphate (pH 7.5 @ 25°C) |
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