| Description |
After 48 hours of transfection, cells overexpressing Myc-PARP1 were harvested. Cells were lysed and immunoprecipitated with anti-Myc antibodies according to co-immunoprecipitation protocol. Then the immunoprecipitated proteins were subjected to Western blot. The bands corresponding to PARP1 were excised in gel digested with trypsin and subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. MS experiments were performed on a nanoscale UHPLC system (EASY-Nlc1000 from Proxeon Biosystems, Odense, Denmark) connected to an Orbitrap Q-Exactive equipped with a nanoelectros pray source (Thermo Fisher Scientific, Bremen, Germany). The raw data were processed using Proteome Discoverer (PD, version 2.1), and MS/MS spectra were searched against the reviewed Swiss-Prot human proteome database. Only peptides with at least six amino acids in length were considered. The peptide and protein identifications were filtered by PD to control the false discovery rate (FDR) < 1%. At least one unique peptide was required for protein identification. |
