returned to 3% B in 2 min and re-equilibrated for 8 minutes, for a total run time of 60 minutes. The Q Exactive Plus was operated in data-dependent mode, with a full MS1 scan followed by 8 data-dependent MS2 scans. The full scan was done over a range of 400–1400 m/z, with a resolution of 70,000 at m/z of 200, an AGC target of 1e6, and a maximum injection time of 50 ms. The MS2 scans were performed at 17,500 resolution, with an AGC target of 1e5 and a maximum injection time of 250 ms. The isolation width was 1.5 m/z, with an offset of 0.3 m/z, and a normalized collision energy of 27 was used.
