Conclusion :
An automated, high-throughput workflow for the enrichment of phosphopeptides from cell lysates was developed. Accurate measurement of phosphorylated proteins and peptides is crucially important due to their central role in cell signaling pathways. Unfortunately, phosphopeptides are typically present at very low levels in cell lysates. An efficient and robust pre-analytical enrichment step for phosphopeptides facilitates the understanding of these fundamental biological pathways.

The enrichment of phosphopeptides described in this report using magnetic TiO2 beads was comprehensive and selective. The described workflow enabled the discovery and identification of phosphopeptides in complex samples with the subsequent development of targeted SRM-based assays that may be useful for clinical research applications.

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