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제목 Milo 등록일 2018-12-11 17:12:32

Milo

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.

Milo - Single-Cell Western - Simplify FACS Studies

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.

 

 

 

 

 

 

 

 

 

 

 

 

 

Milo chart - heterogeneity - 51% KRT8+ cells

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.

Milo chart - Protein validation of my single-cell RNA-seq data

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!

Milo chart - multiplexing

I need to simplify my phospho-flow signaling studies

Eliminate 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.

Milo chart - STAT1 signaling - Eliminate the fixation and permeabilization steps of flow/FACS

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/FACS catalog. Don't get stuck without a flow antibody against your target of interest again.

Milo chart - Milo allows you to use the large commercial catalog of Western antibodies which is 10-100x larger than the flow catalog.

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!

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.

Milo chart - Measure gene editing efficiency or effect in low efficiency gene editing systems

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.

Milo picture - Measure protein expression in low abundance samples and characterize highly enriched FACS-sorted cell populations

Requirements & compatibility

Sample typeSuspension containing >10,000 cells
Cell diameter7-25 µm in suspension
Cell typeMammalian cells; globular in suspension and unfixed
Antibody requirementStandard unlabeled primaries and fluorescent secondaries
Other equipment neededOpen-format fluorescence microarray scanner capable of 5 µm resolution

Performance & specifications

Typical cell dilutionsYield capture and analysis of 1,000-2,000 cells per scWest chip
Molecular weight (MW) range15-175 kDa
MW resolution10% differences in distinct spectral channels, as low as 30% differences in same spectral channel
Typical target multiplexingUp to four proteins per cell by spectral and size-based multiplexing
Twelve-plus proteins per cell using stripping & reprobing
Workflow time4-6 hours

 

 

 

공지사항
Cat No. 제품명
P100 Milo System & Starter Package
P500 Two-color Microarray Scanner without Computer
P700 Two-color Microarray Scanner with Computer
P800 Two-color Microarray Scanner with Autoloader (without computer)
P300 Milo System & Two-color Scanner Package
P900 Two-Color Microarray Scanner with Autoloader and Computer
K800 scWest Calibration Kit
A200 Probing Fixtures
K500 Small scWest Kit
K600 Standard scWest Kit
A300 Three-Plex Antibody Probing Fixtures
R252 5X Wash Buffer
035-003 scWest chip canisters (4 pk)
A100 Electrophoresis Cell
K700 Large scWest Kit
A900 Single-Cell Western Application Support
035-023 Milo Tweezers
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