Additionally, at each time point, many more peptides containing the N-glycan motif (NXT/S) and a deamidation modification are identified in the PNGase F sample compared to the control sample (Fig ). 156 unique potential N-glycosylated peptides were identified across all replicates and timepoints, with an average of 97 ± 25 potential N-glycosylated peptides identified per sample (n = 9). A list of these peptides for each biological replicate can be found in the Supporting Information (SI Tables ). Because PNGase F is used to remove Nglycans from proteins and results in the deamidation of the asparagine residue (a mass increase in 0.98 Da), we hypothesize that peptides containing both the motif and the deamidation shift were successfully deglycosylated with PNGase F. Furthermore, given that the number of total peptides and the number of peptides containing the N-glycan motif and deamidation modification does not improve with increased deglycosylation time, we conclude that trypsin and PNGase F may be added simultaneously to the S-Trap. This allows for the deglycosylation assay to be run in the same amount of time as a standard digestion.