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. Author manuscript; available in PMC: 2017 Nov 9.
Published in final edited form as: Anal Chem. 2015 Sep 30;87(20):10222–10229. doi: 10.1021/acs.analchem.5b02983

Figure 2.

Figure 2

Response curve for a digested 6 protein mix spiked into a complex matrix (C. elegans whole cell lysate). (A) The mixture of six predigested proteins was spiked into digested C. elegans whole cell lysate (1 μg/μL and 1 μg on column) at 8 concentrations from 15 attomol/μL to 62.5 femtomol/μL (0 = blank, 0.015–62.5 fmol/μL, loading 1 μL sample on column). Two replicate concentration curves were acquired on the TripleTOF 5600 (QqTOF in PRM mode, top) and on the QTRAP 5500 (QQQ in SRM mode, bottom), respectively. Peak areas of extracted fragment ion transitions are summed per peptide. Curves are displayed for peptides HLVDEPQNLIK2+ (m/z 653.36) and YSTDVSVDEVK2+ (m/z 621.30), respectively. (B) Peak area CV display for triplicate acquisitions of a digested 6 protein mix (50 fmol/μL) spiked into a complex C. elegans matrix (1 μg/μL) monitoring 15 peptides in PRM mode with 10–24 transitions per peptide (50 fmol analyte injected). Each extracted fragment ion transition is displayed as a vertical colored line. 216 of 240 transitions showed CVs < 20% (Table S-2F, Supporting Information).