Milo
Single-Cell Westerns
Milo™ is the world's first automated single-cell western (scWestern) 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 protein analysis technique.
I need to profile heterogeneity in complex samples using single-cell western analysis
Milo measures protein expression in ~1,000 single-cells in one 4 hour scWestern 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 analyze single-cell proteomics to determine the percentage of cells in your sample that are target positive.

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 using our scWestern instrument Milo. Plus, scWest chips can be archived for up to 9 months after you run them so you have plenty of time to get your RNA sequencing results back before you have to probe for your targets of interest for single-cell proteomics data. Learn how to use single-cell westerns to validate single-cell RNA-seq data.

I need multiplexing to obtain single-cell proteomics data
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! View some of our common scWestern FAQs.

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.

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.

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.

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'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.

Application notes
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 |