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  • FAQ: How much Remove-iT® Endo S should I use to deglycosylate a glycoprotein under native conditions?

    When working with native IgG, we recommend using 1 µL of Remove-iT® Endo S per 100µg of native IgG. When working with other various glycoproteins (such as Fetuin) we recommend using a modified denaturing protocol, detailed below.

    1. Combine 100 µg of glycoprotein, 1 µL of 400mM DTT and H2O (if necessary to make a 10 µL total reaction volume.
    2. Denature glycoprotein by heating reaction at 55°C for 10 minutes.
    3. Make a total reaction volume of 20 µL by adding 2 µL 10X G6 Reaction Buffer, H2O and 1-2 µL Remove-iT® Endo S.
    4. Incubate reaction at 37°C for 1 hour.

    Note: To deglycosylate various glycoproteins, longer incubation times as well as more enzyme may be required. Reactions may be scaled-up linearly to accommodate larger reaction volumes.