Reveal Cell Subtypes, Protein Isoforms, and Phospho-Protein Heterogeneity with Milo

Single-Cell Westerns on Milo™ make measurements that are difficult or impossible to make with conventional Western blots or flow cytometry. Here, we uncover three distinct cell subpopulations that differ based on single- or co-expression of two different protein targets and that could not be distinguished using conventional Western blots or flow cytometry. We also demonstrate Milo's ability to detect discrete cell signaling states by measuring multiple phospho-proteins in individual cells within a population. Finally, we demonstrate how Milo can simultaneously quantify phospho- and total protein expression in each individual cell and quantify how the extent of phosphorylation varies across stimulated and unstimulated cell populations.