At present, the high throughput phosphorylated proteome analysis mainly adopts the method of IMAC affinity chromatography-reversed-phase liquid chromatography-tandem mass spectrometry - database retrieval. The advantage of this method is that each soluble phosphorylated peptide segment, regardless of its length, can be enriched, and the sample eluted by IMAC column can be directly combined with LC for subsequent MOLD-TOF/TOF mass spectrometry identification. The enriched peptide segment can also be further analyzed by two-dimensional acid bump peptide (2D-PP). This method provides important information about protein phosphorylation state that other methods cannot provide.
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