20210901_WXY_TMT_2plex_3Rep_Huvec_proteins

ABSTRACT: Cells were harvested with intra-cellular proteins extracted and digested with trypsin. Peptides were then labeled with TMT using a TMT10plex mass tag labeling kit (Thermo Fisher Scientific) per the manufacturer’s instructions. TMT-labeled peptides were then separated by high pH reverse-phase high performance liquid chromatography (HPLC) with C18 columns (Agilent BioTek) and dried in a vacuum centrifuge. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was subsequently performed. All fragments were sequentially dissolved in aqueous solution (0.1% formic acid and 2% acetonitrile), loaded onto a home-made reverse-phase analytical column, and subjected to an EASY-nLC™ 1000 ultraperformance liquid chromatography (UPLC) system (Thermo Fisher Scientific) at a constant flow rate of 400 nL/minutes. For MS settings, the applied electrospray voltage was 2.0 kV and the m/z range was 350 to 1800 for the complete scan. Peptide fragments were quantified with Parallel Reaction Monitoring (PRM). Proteins were identified by Swissprot database and quantified using Proteome Discoverer 2.0.

ORGANISM(S): Homo Sapiens

SUBMITTER: Xue Chen  

PROVIDER: PXD046601 | iProX | Wed Nov 01 00:00:00 GMT 2023

REPOSITORIES: iProX