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  • FAQ: What are the typical reaction conditions for β-N-Acetylglucosaminidase

    1. Combine 1 µg of glycoprotein or 100nM oligosaccharide and H2O (if necessary) to make a 8 µl total reaction volume.
    2. Adding 1 µl of 10X G1 Reaction Buffer and 1 µl of 10X BSA (diluted 1:10 from 100X concentration) to make a 10 μl total reaction volume.
    3. Incubate reaction at 37°C for 1 hour.

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