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  • Reaction Conditions for Remove-iT® PNGase F (P0706)


    Optimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:

    1. Combine 10–20 µg of glycoprotein, 1 µl of 10X DTT and H20 (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 GlycoBuffer 2, H20 and 1–5 µl Remove-iT PNGase F.
    4. Incubate reaction at 37°C for 1 hour.