Unbiased characterization of the senescence-associated secretome using SILAC-based quantitative proteomics

Approaches based on the combination of mass spectrometry (MS) and quantitative methods have the potential to generate unbiased, thorough proteomic catalogues. In particular, stable isotope labeling with amino acid in cell culture (SILAC) has been used to perform highly accurate quantitative comparisons between the proteomes of different cell lines, treatments, or even animal models. Here, we describe how we have taken advantage of SILAC-based quantitative proteomics and inducible cell systems of oncogene-induced senescence to make an unbiased characterization of the senescence-associated secretome. This approach could be used to analyze the effect of diverse molecules on the senescence secretome or to catalogue unrelated secretomes.