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Experimental design. Control, HIV and HIV-encephalitis human brain lysate samples were reduced, alkylated, and trypsinized separately. Each sample was labeled with a different 8plex iTRAQ reagent and combined. The labeled peptides were enriched for phosphopeptides by TiO2 chromatography. The phosphopeptides were separated on a nanoLC system followed by online high-resolution tandem mass spectrometry.
