Single-cell protein analysis techniques lack resolution, sensitivity or specificity, or require protein tagging. Western blotting avoids these pitfalls but is not amenable to single-cell analysis. Hughes et al. now couple western blotting with single-cell analysis, which enables the simultaneous analysis of ∼2,000 individual cells. Their array-based technique uses a slide coated with polyacrylamide gel and patterned with thousands of microwells, into which a cell suspension is seeded by gravity-driven cell settling, resulting in single-cell occupancy in 40–50% of the wells. Intracellular proteins are then solubilized in the wells, subjected to thin-gel electrophoresis and immobilized. Subsequently, the serial stripping and re-probing of antibodies enables multiplexed analyses of proteins.