FAQ: How do I know whether to follow the “One Step” or the “Two Step” protocol?

Rapid PNGase F has been developed for efficient and fast deglycosylation of antibodies in a simple one step reaction at 50°C. However, some IgGs (i.e. carrying Fab glycosylation) require a pre-denaturing step of 2 minutes at 80°C. We recommend starting with the standard One Step protocol. If there is evidence (i.e. by gel migration or proteomic analysis) that N-glycans remain attached to the protein, follow the Two Step protocol.