Reaction Conditions for Remove-iT® PNGase F (P0706)
ProtocolOptimal incubation times and enzyme concentrations must be determined empirically for a particular substrate. Typical reaction conditions are as follows:
- Combine 10–20 µg of glycoprotein, 1 µl of 10X DTT and H20 (if necessary, to make a 10 µl total reaction volume).
- Denature glycoprotein by heating reaction at 55°C for 10 minutes.
- Make a total reaction volume of 20 µl by adding 2 µl 10X GlycoBuffer 2, H20 and 1–5 µl Remove-iT PNGase F.
- Incubate reaction at 37°C for 1 hour.