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  • Typical Reaction Conditions for α1-3, 6 Galactosidase (P0731)

    • Combine 1 μg of glycoprotein or 100 nM of oligosaccharide and H2O (if necessary) to make a 8 μl total reaction volume.
    • Add 1 μl of 10X G6 Reaction Buffer and 1 μl of 10X BSA (diluted 1:10 from 100X concentration) to make a 10 μl total reaction volume.
    • Add enzyme (see below)
    • Incubate at 37°C for 1 hour.

    Notes
    • The amount of exoglycosidase enzyme required varies when different substrates are used. Start with 1-2 μl for 1ug of glycoprotein or 100 nM of oligosachride for one hour in a 10-25 μl reaction. If there is still undigested material, let the reaction go overnight.
    • The reaction can be scaled up linearly
    • Several exo- and endo-glycosidases can be used together in one digest. For buffer recommendations, please referance the Glycobiology Double Digest Chart .
    • For a more detailed characterization of several glycosidase enzymes, please reference the Detailed Characterization of Several Glycosidase Enzymes page.