Why Single-Cell Westerns?
Decode the mix
Many cell populations, such as tumor cells or differentiating stem cells are heterogeneous. 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. This approach allows simultaneous measurement of multiple proteins in over 1,000 single cells using commercially available Western-validated antibodies.
Identify cell subpopulations you can't see with conventional Westerns
Single-Cell Westerns with Milo reveal cell subpopulations you can't see by conventional Western blotting, which pools thousands or millions of cells to give averaged measurements. Milo can measure protein expression of multiple targets in thousands of individual cells in a single run, so you can identify subpopulations that you just can't measure with traditional Westerns.
Resolve protein isoforms in individual cells
Cell function and behavior is controlled by complex, interconnected signaling pathways and dysregulation of a single component can have potent effects on a myriad of cellular functions. Single-Cell Westerns on Milo provide molecular weight sizing capabilities, which is key to enabling the measurement of different protein isoforms at the single-cell level that are bound by a common antibody. This measurement would be unresolved by other methods, such as flow cytometry.
Identify target expression heterogeneity
Do you want to distinguish between cells that express your target at low levels and cells that express your target at high levels? Milo easily measures these cell-to-cell differences and quantifies variations in the amount of protein present between cells in your sample. For example, you could measure a signaling protein that is upregulated in response to a stimulus and see how variable the response to that treatment is across your cell population.
Count and quantitate at the same time
Each Single-Cell Western uses protein size and antibody binding information for your target of interest to give you a quantitative measure of target abundance in each cell. Quickly get a count of the number of cells that express a particular protein and quantify variations in the amount of protein present in each cell of your sample.
How does Milo let you make measurements you can't make any other way?
Milo 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 four proteins per cell simultaneously.
Read our application note to learn how Milo lets you quantify expression heterogeneity of your target and the percentage of cells in your sample that are target positive.
Read our application note to learn how Milo can measure distinct cell subpopulations that cannot be distinguished with conventional Westerns or by flow cytometry. You'll also learn how he detects discrete cell-signaling states by measuring multiple phospho-proteins in individual cells within a population. Finally, how Milo can simultaneously quantify phospho- and total protein expression in each individual cell and then quantify how the extent of phosphorylation varies across stimulated and unstimulated cell populations is outlined.
Want to learn more about Milo?
Watch the video and see how Milo can automate 1,000 single cell separations in just 4 hours.
If you're curious about pricing, just send us your request.