Quantitative LC/MS/MS was performed using an EvoSep One UPLC coupled to a Thermo Orbitrap Astral high resolution accurate mass tandem mass spectrometer (Thermo). Briefly, each sample loaded EvoTip was eluted onto a 1.5 um EvoSep 150um ID x 15cm performance (EvoSep) column using the SPD30 gradient at 55C. Data collection on the Orbitrap Astral mass spectrometer was performed in a data-independent acquisition (DIA) mode of acquisition with a r=240,000 (m/z 200) full MS scan from m/z 380-1080 in the OT with a target AGC value of 4e5 ions. Fixed DIA windows of 5 m/z from m/z 380 to 1080 DIA MS/MS scans were acquired in the Astral with a target AGC value of 5e4 and max fill time of 8 ms. HCD collision energy setting of 27% was used for all MS2 scans. The total analysis cycle time for each sample injection was approximately 40 min
[doi:10.25345/C56M33H15]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: label free, astral, phosphorylation ; DatasetType:Proteomics
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Principal Investigators: (in alphabetical order) |
Joseph Heitman, MD PhD, Duke University, United States |
| Submitting User: | es3064 |
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