Protein kinase C over-expressed into rat neonatal myocytes were lysed in 8M Urea, 0.1% SDS and trypsin digested. Peptides were fractionated by SPE-SCX and enriched by TiO2. Phosphopeptides were analyzed by LC/MS on a Thermo LTQ Elite Orbitrap. Raw files were searched using Sorcerer-Sequest and post search analysis and processing was done with Scaffold and Scaffold PTM.