Protein phosphorylation controls fundamental biological functions that are often deregulated in disease. Therefore, system-level understanding of complex pathophysiological processes requires methods that can be used to profile and quantify protein phosphorylation as comprehensively as possible. Here we present a detailed protocol to enrich phosphopeptides from total cell lysates in a form amenable to downstream analysis by mass spectrometry. Using these techniques, we have detected several thousands of phosphorylation sites in the NIH-3T3 cell line.
CITATION STYLE
Alcolea, M. P., & Cutillas, P. R. (2010). In-depth analysis of protein phosphorylation by multidimensional ion exchange chromatography and mass spectrometry. Methods in Molecular Biology (Clifton, N.J.), 658, 111–126. https://doi.org/10.1007/978-1-60761-780-8_6
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