Cells: human medulloblastoma Daoy cells

Experiment:  

The medulloblastoma Daoy cell line was obtained from American Type Culture Collection (ATCC). Cells were cultured in improved minimum essential medium (IMEM) (Invitrogen, Carlsbad, CA) 
supplemented with 10% fetal bovine serum (Atlanta Biologicals, Inc., Lawrenceville, GA). Cells were grown until 90% confluence and then harvested to prepare RNA and protein extracts for further 
analyses.

To prepare protein extracts, Daoy cells were re-suspended in an equal volume of lysis buffer (50mM Tris pH 8.0, 50mM NaCl, 1mM EDTA and 1 tablet of Complete Proteinase Inhibitor (Roche)/10ml) 
and incubated on ice for 30 minutes. Cells were then lysed with a dounce homogenizer and the soluble protein extracts obtained after centrifugation.

CYS modification: 
YES

Mass spectrometry:
LC/LC-MS/MS on LCQ

Analysis:

SEQUEST (Bioworks)
Peptide/ProteinProphet v4.0