Fig. 2

Experimental workflow used to identify salivary N-glycosites. Parotid and SMSL salivas collected as the ductal secretions were separated by preparative SDS-PAGE followed by rastering of the gel bands and in-gel trypsin digestion. N-glycopeptides were captured from the peptide extracts via their ability to bind to at least one of four lectins (AAL, JAC, WGA and/or LEA). After elution and PNGase F removal of N-linked glycans, LC–MS/MS was performed to identify the original sites of oligosaccharide attachments and the protein scaffolds