In this project we report 133 biologically distinct mouse liver cell (Hepa 1-6 and FL83B) and mouse tissue (brain, kidney, and liver) phosphoproteomes prepared using a newly developed scalable, single-run phosphoproteomics platform, using titanium dioxide (TiO2) based phosphopeptide enrichment and performed in 96-well format. We combine this technology with in situ hepatic stimulation, to generate time-resolved maps of insulin signaling in the mouse liver.