Normal liver tissue proteome analysis

Liver tissue was homogenized and sonicated in the lysis buffer. Whole cell protein extract was reduced and alkylated and digested by trypsin. <br>Peptide mixture was analyzed on Q Exactive HFX mass spectrometer coupled to UHPLC chromatography system. Two dimensional fractionation of peptides included one more separation on Agilent 1200 Series RP chromatography in alkaline conditions with fractions collected and also analyzed by MS. The fractions were also analyzed by SRM SIS analysis to detect proteins encoded by the 18th human chromosome. The analysis was carried out at 6495 Triple Quad LC/MS system (Agilent). Stable isotope labelled peptides were used as internal standard.The Shotgun data was processed with MaxQuant software v1.6.3.4 by target/decoy method. Cleavage by trypsin, max number of missed cleavages - 2. Oxidation of M and Acetylation of protein N-term were set as variable modifications. Carbamidomethylaion was set as fixed modification.