Proteome of senescent human lung fibroblasts and Panc-1 cells

ABSTRACT: Proteomic analysis of proliferating and senescent normal healthy lung fibroblasts (NHLFs) and Panc-1 cells. Senescence was induced using 5 uM Palbociclib for 6 days. Cells were lysed in SDS lysis buffer (2% SDS in 100 mM TEAB), boiled (15 min, 95 °C, 500 rpm) and sonicated in a sonication bath (10 min). DNA was sheared with Benzonase® Nuclease (Merck Millipore) and 2 mM MgCl2 at 37 °C (30 min, 500 rpm). Reversibly oxidized cysteines were reduced with 10 mM TCEP followed by alkylation with 20 mM IAA (each step 30 min, 22 °C, 600 rpm, in the dark). Proteins were acetone precipitated and the protein pellet was resuspended in 100 µL 100 mM TEAB. Proteins were digested overnight with LysC/trypsin (1:50 protease:protein ratio, 37 °C, 600 rpm) Peptides were desalted by solid-phase extraction using Oasis HLB cartridges (10 mg; Waters), resuspended in 3% ACN/0.1% TFA and analysed on an Orbitrap Eclipse Tribrid mass spectrometer in combination with the FAIMS Pro Interface (Thermo Scientific).

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Karim C. El Kasmi  

PROVIDER: MSV000090607 | MassIVE | Fri Oct 28 04:46:00 BST 2022

SECONDARY ACCESSION(S): PXD037794

REPOSITORIES: MassIVE