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Single-Cell Westerns
Single-Cell your Western with Milo
Protein expression measurements made at the population level often overlook significant differences in expression between individual cells and the existence of cell subpopulations. Milo combines single-cell resolution with the power and versatility of Westerns. Identify target expression heterogeneity, visualize cell subpopulations you can’t see with conventional western blots, simplify phospho-flow studies, and validate your single-cell RNA-seq data at the protein level.
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I need to profile heterogeneity in complex samplesMilo measures protein expression in ~1,000 single-cells in one 4 hour experiment so you can quantify expression heterogeneity of your target and the percentage of cells in your sample that are target positive. |
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I need protein validation of my single-cell RNA-seq dataUse conventional western antibodies to validate your single-cell RNA-seq data with single-cell protein data. Plus, scWest chips can be archived for up to 9 months after you run them so you have plenty of time to get your sequencing results back before you have to probe for your targets of interest. |
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I need multiplexing to detect multiple proteins at the single-cell levelSimultaneously detect 12+ proteins in your sample, all at the single-cell level, using spectral and size-based multiplexing strategies. You can even strip & reprobe scWest chips up to 9 times for higher multiplexed studies! |
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I need to simplify my phospho-flow signaling studiesEliminate the fixation and permeabilization steps of flow/FACS! Milo chemically lyses the cells captured on the scWest chip before analysis to gain access to intracellular and intranuclear compartments more easily than with flow cytometry. By lysing the cells, Milo can detect challenging proteins like transcription factors and even methylated histones! You can also use Milo's sizing step to separate out non-specific binding, which is more common with phospho-specific antibodies and cannot be resolved with flow. |
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I need to detect proteins that lack good flow antibodiesMilo is an open platform so you can use the large commercial catalog of Western antibodies which is 10-100x larger than the flow/FACS catalog. Don't get stuck without a flow antibody against your target of interest again. |
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I want to measure protein isoform heterogeneityMilo's molecular weight sizing step can resolve protein isoforms that differ in molecular weight, allowing you to measure how many cells in your sample express one isoform, the other isoform, or both isoforms. Try that with flow! |
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I need to measure gene editing efficiency or effect in low efficiency gene editing systemsMilo can quantify the percentage of cells in your sample that express an inserted gene, even in low efficiency systems. Plus, you can simultaneously measure your edited gene and downstream effect markers to understand the impact of the gene on the cells that have been edited. |
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I need to measure protein expression in low abundance samplesMilo can analyze samples which contain as low as 10,000 single-cells so you no longer have to have collect millions of cells to analyze protein expression. The number of cells captured scales with the number of cells loaded so even lower abundance samples are possible. Milo can also characterize highly enriched FACS-sorted cell populations which don't contain enough cells to analyze with other techniques. |
More Information About Milo
| Feature | Description |
|---|---|
| Requirements & Compatibility | |
| Sample type | Suspension containing >10,000 cells |
| Cell diameter | 7-25 µm in suspension |
| Cell type | Mammalian cells; globular in suspension and unfixed |
| Antibody requirement | Standard unlabeled primaries and fluorescent secondaries |
| Other equipment needed | Open-format fluorescence microarray scanner capable of 5 µm resolution |
| Performance & Specifications | |
| Typical cell dilutions | Yield capture and analysis of 1,000-2,000 cells per scWest chip |
| Molecular weight (MW) range | 15-175 kDa |
| MW resolution | 10% differences in distinct spectral channels, as low as 30% differences in same spectral channel |
| Typical target multiplexing | Up to four proteins per cell by spectral and size-based multiplexing Twelve - plus proteins per cell using stripping & reprobing |
| Workflow time | 4-6 hours |
