NEB, P0769S, α1-3,4 Fucosidase
Catalog Number: P0769S
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| <b>Unit Definition</b> One unit is defined as the amount of enzyme required to cleave > 95% of the α-fucose from 1 nmol of Galβ1-4GlcNAcβ1-3(Fucα1-3)Galβ1- 4Glc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 μl <b>Reaction Conditions</b> 1X GlycoBuffer 1 Supplement with 100 µg/ml Recombinant Albumin (Low-glycerol) 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 @ 25°C <b>Heat Inactivation</b> 65°C for 10 minutes <b>Molecular Weight</b> Apparent | 56 kDa <b>Unit Assay Conditions</b> Two fold dilutions of a1-3,4 Fucosidase are incubated with 1 nmol AMC-labeled substrate in 1X GlycoBuffer 1 supplemented with 1X rHSA 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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