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Optimization of TiO2 phosphopeptide enrichment using 0.5 mg of rat brain lysate and various additives to reduce the binding of non-phosphopeptides. Peptide-to-bead ratio is 1:4 (w/w). (A) The number and percentage of identified phosphopeptides without and with three additives:1 mM phosphate, saturated glutamic acid and 2 M lactic acid, respectively. The isolated peptides were analyzed by 1 h LC-MS/MS. (B) The effect of titrated levels of phosphate on phosphopeptide enrichment.
