Tryptic peptides were separated on a nanoACQUITY UPLC analytical column (BEH130 C18, 1.7 μm, 75 μm x 200 mm, Waters) over a 165-minute linear acetonitrile gradient (3 – 40%) with 0.1 % formic acid on a Waters nano-ACQUITY UPLC system and analyzed on a coupled Thermo Scientific Orbitrap Fusion Lumos Tribrid mass spectrometer as previously described (Williamson et al. 2016 (link); Defnet et al. 2019 (link); Kim et al. 2019 (link)). Full scans were acquired at a resolution of 240,000, and precursors were selected for fragmentation by collision induced dissociation (normalized collision energy at 35 %) for a maximum 3-second cycle.
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Liquid Chromatography-Mass Spectrometry Proteomics
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Corresponding Organization : University of Maryland, Baltimore
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Variable analysis
independent variables
- Acetonitrile gradient (3 - 40%)
- Tryptic peptide separation on nanoACQUITY UPLC analytical column
- Mass spectrometric analysis on Thermo Scientific Orbitrap Fusion Lumos Tribrid mass spectrometer
- NanoACQUITY UPLC analytical column (BEH130 C18, 1.7 μm, 75 μm x 200 mm, Waters)
- 0.1% formic acid
- Full scans at a resolution of 240,000
- Collision induced dissociation (normalized collision energy at 35%) for a maximum 3-second cycle
- Positive controls not explicitly mentioned
- Negative controls not explicitly mentioned
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