Milo

Single-Cell Westerns

Milo™ is the world's first automated single-cell western platform. The instrument measures protein expression in thousands of cells in a single run, allowing you to profile heterogeneity in your samples through single-cell analysis. 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 blot antibodies to measure ~12 proteins per cell using a variety of multiplexing strategies. Our Single-Cell Western technology on Milo unlocks the single-cell proteome to measure more of the proteome than was possible with any other single-cell technique.

Use Milo to Perform Automated Single-Cell Western, Simplify FACS Studies, Measure Profile Heterogeneity, and Validate RNA-Seq Data

How Can Milo Help You?

I need to profile heterogeneity in complex samples using single-cell analysis

Milo measures protein expression in ~1,000 single-cells in one 4 hour single-cell western experiment so you can quantify heterogeneity of your target at the single-cell level to determine expression patterns. Using antibodies that work for traditional western blot, Milo provides a platform to perform protein expression at the single-cell level to determine the percentage of cells in your sample that are target positive.

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Milo Automated Single-Cell Western Measurement Heterogeneity Chart of 51% KRT8+ Cellular Expression

I need protein validation of my single-cell RNA-seq data

Use conventional western blot 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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Using Protein Validation with Single-Cell RNA-Seq Data on Milo, the Automated Single-Cell Western Instrument

I need multiplexing to detect multiple proteins at the single-cell level

Simultaneously 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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Multiplexing Proteins on Milo, the Automated Single-Cell Western Instrument Allows 12+ protein detection per sample

I need to simplify my phospho-flow signaling studies

Eliminate the fixation and permeabilization steps of flow cytometry! 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 cytometry methods.

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Milo Can Analyze Data Like Phosphorylation States as Depicted in pSTAT1 Signaling Graphs. Milo Eliminates the Need For Fixation and Permeabilization Steps of Flow Cytometry using ProteinSimple's Automated Single-Cell Western

I need to detect proteins that lack good flow cytometry antibodies

Milo is an open, automated single-cell western platform where you can use the large commercial catalog of western blot validated antibodies, which is 10-100x larger than the flow cytometry antibody catalog. Don't get stuck without a flow antibody against your target of interest again. Use Milo's Single-Cell Western technology to perform single-cell protein analysis.

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Milo Measurement Charts of H3K27me3 Antibody, Histone H3 Antibody, and FoxP3 Antibody Showing Target Signal Strength and Counts - Milo Works with Certified Simple Western Western Blot Antibodies

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. This capability is made possible by Milo's Single-Cell Western technology which surpasses other methods like flow cytometry.

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Milo chart - Measure protein isoform heterogeneity - 2 isoforms resolved by molecular weight

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.

Measure Gene Editing Efficiency with Milo's Single-Cell Western Technology by ProteinSimple

I need to measure protein expression in low abundance samples

Milo's automated single-cell western tests can analyze samples which contain as low as 10,000 single-cells so you no longer have to have to 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.

Use Milo's Automated Single-Cell Western Capabilities to Measure Protein Expression in Low Abundance Samples and Characterize Highly Enriched FACS-Sorted Cell Populations
Single-Cell Applications

 
Cancer

Simple Western Instrument Cancer Research Applications

 
SARS-CoV-2

Simple Western Instrument SARS-CoV-2 Research Applications

Cell & Gene Therapy

Simple Western Instrument Cell & Gene Therapy Research Applications

Targeted Degradation

Targeted Degradation

 

SARS-CoV-2 Serology

SARS-CoV-2 Multi-Antigen Serology Module
 
 
 
Application Notes

Application notes

Wes and Milo Synergize to Profile Immune Cell Populations in the Tumor microenvironment- Application Note   
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Mending the Myocardium: Tracking Cardiomyocyte Differentiation with Single-Cell Westerns   
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Wes and Milo Tumor Microenvironment App Note Japanese   
Wes and Milo Synergize to Profile Immune Cell Populations in the Tumor Microenvironment   
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Identify and Quantify Neural Subtypes with Single-Cell Westerns Application Note   
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Dissociation of Mouse Neural Tissue for Single-Cell Western Analysis Application Note   
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Multiplexed Single-Cell Western Analysis of Red Blood Cells for Biomarker Detection in Blood and Neurodegenerative Disorders   
Going with The Flow: Using Milo to Streamline Your Flow Cytometry Experiments   
Generating Single-Cell Western Protein Heterogeneity Data with an InnoScan 710 Scanner   
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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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Specifications

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