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Experimental procedure for quantitative analysis of secreted glycoproteins using a boosting channel. The cells are prelabeled with Ac4GalNAz for glycoprotein enrichment in either SCM (quantification channels) or SFM (boosting channel). The media are collected and glycoproteins are tagged with DBCO-sulfo-biotin. The enriched glycopeptides are labeled with the TMT 6-plex reagents, deglycosylated with PNGase F, and analyzed with LC-MS/MS.
