1. Standard concentration curves for stable isotope-labeled and acetyllysine containing peptides.
Standard concentration curves for stable isotope-labeled and acetyllysine containing peptides.A and B, YAPVAKacDLASR (A, succinate dehydrogenase complex, subunit A) and LVSSVSDLPKacR (B, 3-hydroxy-3-methylglutaryl-CoA synthase 2) spanning a concentration range from 4 attomoles to 25 femtomoles. Peptides were diluted into either a simple matrix (red triangles, 25 fmol “six protein mix”) or a complex matrix (blue circles, mitochondrial lysate from mouse liver, 0.3 μg on column), and acquired on a TripleTOF 5600 mass spectrometer in triplicates. MS1 filtered peak area curves for precursor ions M with m/z (A, heavy YAPVAKacDLASR) and m/z (B, heavy LVSSVSDLPKacR). Weighted regression lines and calculated LOQ are indicated (also see supplemental Table S3), and weighted linear regression slopes were determined as 1.03 and 1.03 for YAP and 0.99 and 1.00 for LVS peptides in the different matrices. C, peak area CVs across three replicates for each concentration point (0.037–25 fmol) in complex matrix are shown for six targeted acetyllysine peptides with 20 and 30% CV cutoffs indicated (also see supplemental Figs. S6, C and F). D, high throughput analysis showing peak area CVs for 366 peptides across 27 independently acquired runs in a complex mitochondrial lysate background matrix. Peak area CVs are displayed against MS1 filtered precursor m/z.
Birgit Schilling et al. Mol Cell Proteomics 2012;11:
© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.