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. Author manuscript; available in PMC: 2018 Nov 21.
Published in final edited form as: Biochem J. 2017 Nov 21;474(23):3963–3984. doi: 10.1042/BCJ20170615

Figure 4.

Figure 4

Schematic of sample preparation for SILAC quantification and representative example of quantification by LC-MS/MS. Cells grown in media supplemented with either light or heavy lysine and arginine were transfected with DCBLD(X)–Flag in kinase inactive/inhibited (light) or active (heavy) conditions. Post-IP, heavy and light immune complex pairs were combined and analyzed via LC-MS/MS to determine relative abundance of phosphopeptide ions between each state. Heavy-to-light ratios (H:L) of monoisotopic peak intensities were normalized to DCBLD(X) peptides that were found not to be modified (loading control).