Computational protocol: Proteomic analysis of the SIRT6 interactome: novel links to genome maintenance and cellular stress signaling

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Protocol publication

[…] After heat-shock, MEF cells were harvested after 1 h30 or recovered for 24 h at 37C before harvesting. Cells were rinced and resuspended in ice cold Phosphate Buffered Saline (PBS) containing propidium iodide and immediately analyzed by flow cytometry. Flow cytometry was performed on a BD-LSRII cytometer (BD-Bioscience) at the Stanford Shared FACS Facility (SSFF). For detection of propidium iodide staining, sample were excited with a 532 nm Green laser using a 640 a Low-Pass splitter and 670/14 Low-Pass filter. For each sample, 10000 events were collected. Data were analyzed using the flowjo software (Treestar, Inc). Cells stained with propidium iodide were gated (see ) and their percentage compared to the whole population quantified. […]

Pipeline specifications

Software tools EMBOSS, FlowJo
Application Flow cytometry