Figure 6. In-gel deglycolation by PNGase F or Png1p-ΔH1 digestion.

PNGase F or Png1p-ΔH1 was incubated with RNase B in gel in 30 µl of 20 mM pH 7.0 NaHCO₃. All proteins used in the assay were purified at the same time, following the same protocol. The molar ratio of enzyme to substrate was 1∶30 in each reaction. After enzymatic digestion, samples were analyzed by SDS–PAGE followed by Coomassie staining. +CHO represents the glycosylated form of protein, and -CHO represents the deglycosylated form of protein.