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  • FAQ: What is a typical reaction protocol for Remove-iT® Endo D?

    We recommend using 1 µL of Remove-iT® Endo D per 20µg of paucimannose N-linked glycans.
    1. Combine 10-20 µg of glycoprotein, 1 µL of 10X DTT and H2O (if necessary to make a 10 µL total reaction volume.
    2. Denature glycoprotein by heating reaction at 95°C for 3 minutes.
    3. Make a total reaction volume of 20 µL by adding 2 µL 10X G7 Reaction Buffer, H2O and 1 µL Remove-iT® Endo D.
    4. Incubate reaction at 37°C for 1 hour.
    Note: To deglycosylate various glycoproteins under native conditions, longer incubation times as well as more enzyme may be required. Reactions may be scaled-up linearly to accommodate larger reaction volumes.