| <b>Unit Definition</b> One unit is defined as the amount of enzyme required to cleave > 95% of the fucose from 1 nmol of GlcNAc(Fucα1-6)β1-4GlcNAc-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> 20 mM Tris-HCl 50 mM NaCl 1 mM EDTA pH 7.5 @ 20°C <b>Heat Inactivation</b> 65°C for 10 minutes <b>Molecular Weight</b> Apparent | 49 kDa <b>Unit Assay Conditions</b> Two fold dilutions of α1-2,4,6 Fucosidase O are incubated with 1 nmol AMC-labelled 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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