By removing glycans from peptides, either chemically or enzymatically, before or after proteolytic digestion, the base peptides can be identified using traditional bottom-up proteomics without the variable mass addition. A common N-glycosidase is peptide:N-glycosidase F (PNGase F), which is efficient for removal of N-glycans from proteins. PNGase F cleaves the internal glycoside bond between the asparagine residue and the innermost monosaccharide of the N-glycan, N-acetylglucosamine (GlcNAc). It is a widely applicable enzyme, able to hydrolyze high-mannose, hybrid, and bi-, tri-, and tetra-antennary oligosaccharides. PNGase deglycosylation results in a free, complete sugar chain and the conversion of asparagine to aspartic acid through a deamidation modification, that can be easily detected using mass spectrometry by a characteristic mass increase of 0.984 Da at the site of deamidation. Although deglycosylating samples prior to traditional digestion can increase proteomic coverage, the required reactions and sample clean-up can add up to a day to already lengthy sample preparation workflows.