| <b>Unit Definition</b> 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-AMC, in 1 hour at 37°C in a total reaction volume of 10 μl. <b>Reaction Conditions</b> 1X GlycoBuffer 1 Incubate at 37°C 1X GlycoBuffer 1 5 mM CaCl2 50 mM sodium acetate (pH 5.5 @ 25°C) <b>Storage Buffer</b> 50 mM NaCl 20 mM Tris-HCl 1 mM EDTA pH 7.5 @ 25°C <b>Heat Inactivation</b> 65°C for 10 minutes <b>Molecular Weight</b> Apparent | 100 kDa <b>Specific Activity</b> 316,000 units/mg <b>Unit Assay Conditions</b> Two fold dilutions of α2- 3,6,8,9 Neuraminidase A are incubated with 1 nmol AMC-labeled substrate and 1X GlycoBuffer 1 in a 10 μl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via thin layer chromatography (2). |
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