How can I test whether the Rapid PNGase F reaction is complete in 10 minutes?

For a monoclonal antibody, a SDS-PAGE shift can be observed after removal of N-glycans. Typically, this is only a modest change, so it is crucial to run a control along with the real sample to compare the position on a gel, and determine whether deglycosylation is partial or complete (refer to the “Analysis of a Rapid PNGase F Deglycosylation Reaction by SDS-PAGE” protocol). To detect whether or not minor amounts of glycosylated antibody remain, a more sensitive assay is required. For instance, a proteomic analysis by LS-ESI-TOF will show the different glycosylated and non-glycosylated forms present (refer to the “Intact Protein LS-ESI-TOF Protocol”).

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