Sample processing for proteomics
Seven treatments were chosen to capture states near the critical salinity threshold. Eight samples from each treatment were taken from the short-term 85g/kg and 105g/kg treatments and corresponding FW control, and from the long-term 75g/kg treatment and FW control. Six replicates were sampled from the acclimation to LER at 85g/kg and 105g/kg. Extraction, in-solution trypsin digestion of proteins, and peptide separation using a nanoAcquity UPLC system equipped with Symmetry trap and 250 mm x 1.7µm particle size BEH C18 columns were performed as previously reported [29].