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Re-suspended peptides were loaded onto a nanoViper C18 Acclaim PepMap 100 trap column (PN 164946, Thermo Scientific) and resolved using an increasing gradient of ACN in 0.1% Formic acid through a 50 cm 75 um ID EasySpray analytical column (PN ES803A, Thermo Scientific) at a flow rate of 200 nL/min during the first 160 min and 300 nL/min over the last 20 min