FAQ: Should I use the one- or two-step Rapid PNGase F deglycosylation method for my antibody?

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 (e.g., 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 (e.g., by gel migration or proteomic analysis) that N-glycans remain attached to the protein, follow the two-step protocol.