Single-Cell Westerns

Milo™ is the world's first Single-Cell Western platform. He measures protein expression in thousands of single cells in a single run so you can profile heterogeneity in your samples. Just load your cell suspension and the scWest chip captures ~1,000 single-cells. Milo then does a fast, 1 minute SDS-PAGE separation on each single-cell lysate on-chip. Then just probe with your favorite conventional Western antibodies to measure up to 4 proteins per cell simultaneously. Use Single-Cell Westerns to unlock the single-cell proteome and measure more of the proteome than is possible with any other single-cell technique.


How Can Milo Help You?

I need to profile heterogeneity in complex samples

Milo 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 to simplify my phospho-flow signaling studies

Eliminate the fixation and permeabilization steps of flow! 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.

I need to detect proteins that lack good flow antibodies

Milo is an open platform so you can use the large commercial catalog of Western antibodies which is 10-100x larger than the flow catalog. Don't get stuck without a flow antibody against your target of interest again.

I want to measure protein isoform heterogeneity

Milo'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 protein validation of my single-cell RNA-seq data

Use 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.

I need to measure gene editing efficiency or effect in low efficiency gene editing systems

Milo 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.

I need to measure protein expression in low abundance samples

Milo 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.


Application notes

Using Single-Cell Westerns to Validate Single-Cell RNA-Seq Data   
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Reveal Cell Subtypes, Protein Isoforms, and Phospho-Protein Heterogeneity with Milo   
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Adapting the Single-Cell Western Protocol to Detect Histone Modifications    
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Protein Expression Heterogeneity with Milo, the First Single-Cell Western System   
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The Single-Cell Western has Arrived   
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Requirements & compatibility

Sample type Suspension containing >10,000 cells
Cell diameter 7-25 µm
Cell type Mammalian cells; globular in suspension and unfixed
Antibody requirement Standard, commercial unlabeled primaries and fluorescent secondaries
Other equipment needed Open-format fluorescence microarray scanner capable of 10 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
Workflow time 4-6 hours