To further investigate the compatibility of PNGase F with S-Traps, the performance time of PNGase F was examined. Five samples of RNase B were treated with PNGase F and incubated for 1, 5, 15, 60, or 120 minutes, followed by heat shock to stop the reaction. A similar gel shift is observed for all samples, indicating that the deglycosylation reaction reaches completion by the first time point (Fig ). We hypothesize that because PNGase F works quickly and performs the same at the beginning and end of a two-hour period, it is not necessary to delay the addition of trypsin after the addition of PNGase F.