NEB, P0773S, Endoglycoceramidase I (EGCase I)
Catalog Number: P0773S
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Category
TagsGlycobiology
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| <b>Unit Definition</b> One unit of R. triatomea EGCase I was defined as the amount of enzyme required to hydrolyze 1 μmol of ganglioside GM1a per minute at 37°C. <b>Reaction Conditions</b> 1X EGCase I Reaction Buffer Incubate at 37°C 1X EGCase I Reaction Buffer 50 mM sodium acetate 0.1% Triton® X-100 (pH 5.2 @ 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 20 minutes <b>Molecular Weight</b> Apparent | 50 kDa <b>Unit Assay Conditions</b> Two fold dilutions of EGCase I are incubated with 10 nmoles of GM1a substrate in 1X EGCase I Reaction Buffer in a 10 μl reaction. The reaction mix is incubated for 5 minutes at 37°C. Released glycans are labeled with 2-AB, and reaction products are analyzed by ultra-performance hydrophilic interaction liquid chromatography with fluorescence detection. |
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