Glycosylations

[rj@ucsf]

I have a protein with four glycosylation sites, 3 N-linked and 1 O-linked. I have tried to remove the N-linked residues with PNGase F and Endo H without success. Does anyone know of better enzymes or better methods to deglycosylate native proteins?

[Jyona]

Hey actually my protein has 2 glycosylation sites , one of which we mutated and the other one we are trying to digest it with PNGase F and Endo H but I don't know the result right now.......what exact procedure and unit you use to see the enzyme activity. I know PNGaseF is a kind of control but Endo H deglycosylate the protein to some extent.