Protein N-linked glycosylation plays a key role in various biological and pathological processes in the brain. Interestingly, different brain cells such as neurons and astrocytes can have distinct N-glycosylation patterns. However, it remains challenging to analyze brain N-glycosylation in a cell-type specific manner. Here we report the glycoproteomic studies of N-linked glycosylation in primary rat neurons and astrocytes. By combining primary brain cell culture and mass spectrometry-based analysis of intact glycopepties, we identified 6461 glycopeptides, 1684 glycosides and 770 glycoproteins, including 144 sialic acid glycoproteins. These N-linked glycosylated proteins are highly enriched in cell communication and adhesion. Profiling of intact glycopeptides provides a powerful tool for investing the biological function of brain glycosylation.