MassIVE MSV000095110

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Presynaptic Rac1 Interactome in vivo BioID Soderling and Kim

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Description

Quantitative LC/MS/MS was performed on 2 uL of each sample, using a Vanquish Neo UPLC system (Thermo) coupled to a Thermo Orbitrap Astral high resolution accurate mass tandem mass spectrometer (Thermo). Briefly, the sample was first trapped on a Symmetry C18 20 mm x 180 um trapping column (5 ul/min at 99.9/0.1 v/v water/acetonitrile), after which the analytical separation was performed using a 1.5 um EvoSep 150um ID x 8cm performance (EveoSep) column with a 30 min gradient of 5 to 30% acetonitrile with 0.1% formic acid at a flow rate of 500 nanoliters/minute (nL/min) with a column temperature of 50C. 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 380-980 with a target AGC value of 4e5 ions. Fixed DIA windows of 4 m/z from m/z 380 to 980 DIA MS/MS scans were acquired in the Astral with a target AGC value of 5e4 and max fill time of 6 ms. HCD collision energy setting of 27% was used for all MS2 scans.The total analysis cycle time for each sample injection was approximately 35 min. [doi:10.25345/C50C4SW7C] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: bioID, astral, label free

Contact

Principal Investigators:
(in alphabetical order) 		Scott Soderling, Duke University, Dept of Cell Biology, US
Submitting User: es3064
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