The methods of protein extraction and trypsin digestion were performed according to the protocol reported in Sun et al. (2017) (link) with slight modifications. The gradient was comprised of an increase from 6 to 22% of solvent B (0.1% FA in 98% ACN) for 40 min, from 22 to 35% over 12 min, climbing to 80% over 4 min, and then holding at 80% for the last 4 min. A constant flow rate of 300 nL/min was maintained on an EASY-nLC 1000 UPLC system, and the resulting peptides were analyzed by a Q ExactiveTM plus hybrid quadrupole-Orbitrap mass spectrometer (Thermo Fisher Scientific).
The peptides were subjected to Nanospray ionization (NSI) source followed by tandem mass spectrometry (MS/MS) in Q ExactiveTM plus (Thermo) coupled with an online Ultra Performance Liquid Chromatography (UPLC). For MS scans, the m/z scan range was 350 to 1800. Fixed first mass was set as 100 m/z.
The peptides were subjected to Nanospray ionization (NSI) source followed by tandem mass spectrometry (MS/MS) in Q ExactiveTM plus (Thermo) coupled with an online Ultra Performance Liquid Chromatography (UPLC). For MS scans, the m/z scan range was 350 to 1800. Fixed first mass was set as 100 m/z.
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