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.
References
Hughes, A. J. et al. Single-cell western blotting. Nature Methods http://dx.doi.org/10.1038/nmeth.2992 (2014)
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Zlotorynski, E. Single-cell western blotting. Nat Rev Mol Cell Biol 15, 429 (2014). https://doi.org/10.1038/nrm3833
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DOI: https://doi.org/10.1038/nrm3833