| <b>Unit Definition</b> One unit is defined as the amount of enzyme required to cleave > 95% of the terminal, non-reducing β- N -Acetylglucosamine from 1 nmol GlcNAcβ1-4GlcNAcβ1-4GlcNAc-7-amino-4-methylcoumarin (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> No <b>Molecular Weight</b> Apparent | 125 kDa <b>Unit Assay Conditions</b> Two fold dilutions of β- N -Acetylglucosaminidase S 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 (1). |
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