Methods: Ga3+-IMAC magnetic beads were used to capture phosphopeptides from a model system based on b-casein, and then on an enzymatic digestion of a human A431 cell lysate. The peptides were released from the beads and analyzed by reversed phase C18 LC/MS/MS with an
LCQ Deca XP Plus ion trap mass spectrometer.

Results: We present a fast, simple protocol for enriching phosphopeptides in complex mixture using magnetic bead-based IMAC prior to LC/MSn analysis in an ion trap mass spectrometer. Beginning with 100 µg of cell lysate digest, this procedure was used to successfully enrich and identify more than a dozen phosphoproteins from A431 cells.

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