S-nitrosothiol Resin Assisted Capture(SNORAC) coupled with mass spectrometry analysis has been used to study S-nitrosylated proteins in rat cortical neurons. Proteins were captured on the resin by covalently attaching their former S-NO moiety. Then, nistrosylated proteins were eluted by on-bead digestion. Formerly S-nitrosylated peptides were also eluted from the beads in a second step, and analyzed separately for assessment of the nitrosylation site (qualitative analysis). Out of over 600 nitrosyltated proteins, 131 proteins have never been shown to be S-nitrosylated in any system and 612 are new targets of S-nitrosylation in cortical neurons. The site(s) of Snitrosylation were also identified for 59% of the targets.