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  • FAQ: What are the typical reaction conditions for PNGase F?

    Typical reaction conditions are as follows:

    1. Combine 1-20 µg of glycoprotein, 1 µl of 10X Glycoprotein Denaturing Buffer and H2O (if necessary) to make a 10 µl total reaction volume.
    2. Denature glycoprotein by heating reaction at 100°C for 10 minutes.
    3. Make a total reaction volume of 20 µl by adding 2 µl 10X GlycoBuffer 2, 2 µl 10% NP40, H2O and 1-2 µl PNGaseF.
    4. Incubate reaction at 37°C for 1 hour. 

    Note: We recommend limiting PNGaseF to 1/10 (or less) of the total reaction volume to keep final glycerol concentration equal to (or less than) 5%. Reaction may be scaled-up linearly to accommodate large amounts of PNGaseF and larger reaction volumes.

    Links to this resource

    PNGase F