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Quantification of phosphorylation on Thr-66 on eIF2γ. (A) Precursor ion mass scan of the [M + 2H]2+ ion is shown; (B) MS/MS spectra of m/z ion 582.9 illustrating indicative b- and y-ions for peptide spanning residues 60-68 of eIF2γ; and (C) Zoomed in view of the m/z region containing the TMT (tandem mass tags) reporter ions. Calculation of the reporter ion ratio reveals a phosphorylation level of 70.5% for Thr-66.
