PMC:7321036 / 82712-85162 JSONTXT 8 Projects

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Id Subject Object Predicate Lexical cue
T752 0-274 Sentence denotes Digested samples were analyzed on an Orbitrap Exploris 480 mass spectrometry system (Thermo Fisher Scientific) equipped with an Easy nLC 1200 ultra-high pressure liquid chromatography system (Thermo Fisher Scientific) interfaced via a Nanospray Flex nanoelectrospray source.
T753 275-402 Sentence denotes For all analyses, samples were injected on a C18 reverse phase column (25 cm x 75 μm packed with ReprosilPur 1.9 μm particles).
T754 403-488 Sentence denotes Mobile phase A consisted of 0.1% FA, and mobile phase B consisted of 0.1% FA/80% ACN.
T755 489-699 Sentence denotes Peptides were separated by an organic gradient from 5% to 30% mobile phase B over 112 minutes followed by an increase to 58% B over 12 minutes, then held at 90% B for 16 minutes at a flow rate of 350 nL/minute.
T756 700-765 Sentence denotes Analytical columns were equilibrated with 6 μL of mobile phase A.
T757 766-885 Sentence denotes To build a spectral library, one sample from each set of biological replicates was acquired in a data dependent manner.
T758 886-1144 Sentence denotes Data dependent analysis (DDA) was performed by acquiring a full scan over a m/z range of 400-1000 in the Orbitrap at 60,000 resolving power (@200 m/z) with a normalized AGC target of 300%, an RF lens setting of 40%, and a maximum ion injection time of 60 ms.
T759 1145-1224 Sentence denotes Dynamic exclusion was set to 60 seconds, with a 10 ppm exclusion width setting.
T760 1225-1377 Sentence denotes Peptides with charge states 2-6 were selected for MS/MS interrogation using higher energy collisional dissociation (HCD), with 20 MS/MS scans per cycle.
T761 1378-1625 Sentence denotes For phosphopeptide enriched samples, MS/MS scans were analyzed in the Orbitrap using isolation width of 1.3 m/z, normalized HCD collision energy of 30%, normalized AGC of 200% at a resolving power of 30,000 with a 54 ms maximum ion injection time.
T762 1626-1808 Sentence denotes Similar settings were used for data dependent analysis of samples used to determine protein abundance, with an MS/MS resolving power of 15,000 and a 22 ms maximum ion injection time.
T763 1809-1870 Sentence denotes Data-independent analysis (DIA) was performed on all samples.
T764 1871-2169 Sentence denotes An MS scan at 60,000 resolving power over a scan range of 390-1010 m/z, a normalized AGC target of 300%, an RF lens setting of 40%, and a maximum injection time of 60 ms was acquired, followed by DIA scans using 8 m/z isolation windows over 400-1000 m/z at a normalized HCD collision energy of 27%.
T765 2170-2198 Sentence denotes Loop control was set to All.
T766 2199-2332 Sentence denotes For phosphopeptide enriched samples, data were collected using a resolving power of 30,000 and a maximum ion injection time of 54 ms.
T767 2333-2450 Sentence denotes Protein abundance samples were collected using a resolving power of 15,000 and a maximum ion injection time of 22 ms.