FAQ: How much PNGase F, Recombinant should I use to remove my carbohydrate under native or DTT denaturing conditions?

When the protein is not denatured with SDS, PNGase F, Recombinant has to work harder to reach the cleavage site of the carbohydrate (because of the secondary and tertiary protein structure). Sometimes extra enzyme and extended incubation times can help but these values are specific to each protein and must be determined empirically.