Need Protein Validation of Your Single-cell RNA Sequencing (RNA-seq) Data?
Do you have a list of targets from your Single-cell RNA sequencing (RNA-seq) experiments and want to verify protein expression levels for your targets? Only Single-cell Westerns offer the versatility to validate diverse protein targets discovered in your sequencing runs.
Learn About Single-cell Westerns on Milo and Single-Cell RNA Data
With Milo from ProteinSimple, you can perform Single-cell Westerns and measure protein expression in thousands of individual cells in a single run. Good for validating single-cell RNA data at the protein level.
Since mRNA levels do not always correlate with functional protein levels, pairing single-cell RNA data with single-cell protein expression data gives you more accurate and complete conclusions about cellular function.
In this sample, all cells are positive for HIF-1α RNA, but Milo shows that only about half of the cells contain HIF-1α protein.
Milo users are uncovering novel insights at the single-cell level that are leading the publication in the top scientific journals.
How Protein Validation with Single-Cell RNA-Seq Data Works
Here's how protein validation with single-cell RNA-seq data works.
You put your cell suspension on the scWest chip and put the chip in Milo. Milo captures over a thousand cells, lyses them, runs an SDS-PAGE separation on every single-cell lysate, and immobilizes all your separated proteins in just five minutes.
Next, you prep your protein targets on chip with standard primary and fluorescent secondary antibodies and then image chip fluorescence.
Data analysis is then performed for you automatically using ProteinSimple’s Scout software. The entire process from sample loading to data analysis takes just four to six hours with no overnight transfer step.
You can use off-the-shelf primary Western antibodies, so you can be sure you'll find an antibody against all your targets of interest.
Hear From Scientists Using Milo for Gut-Brain Axis Research
Here's what Dr. Maya Kaelberer, a postdoctoral fellow at Duke University, had to say about her experience using Milo to uncover new insights she published in Science about how the gut and brain communicate. Learn more in the Guts and Glory: Validating a Neuroepithelial Circuit using Milo from your peers story.
“In our lab, the way that we've been using Milo is to pair it with kind of single-cell sequencing data so we have the RNA. We know the transcripts are there, but this is actually a way for us to measure and quantify the amount of protein that's in each individual cell. My favorite feature of Milo is definitely how easy it is to use. You basically just put your samples on their slides and pop it into Milo. You run the Gel through for a short period of time, right it's just seconds and having that automated as opposed to manually trying to turn on and off things has made it a lot easier. The power of it, which I really like, is the fact that you can assay a number of cells, in these kind of microwells, and then get their protein and output. So, the software was very easy to use. It’s fairly intuitive which is pretty much what you would want from your software, and it tells you exactly what you need to know. I would recommend Milo to my colleagues, and I have actually discussed it with some of them. We're moving forward with working with it on our NASH study.”
To request quote and learn more about Milo, visit the Milo instrument page.
VALIDATE YOUR SINGLE-CELL RNA-SEQ DATA WITH MILO
Milo provides single-cell protein expression information to validate your single-cell RNA data. Since mRNA levels do not always correlate with functional protein levels, pairing single-cell RNA data with single-cell protein expression data is critical to making accurate and complete conclusions about cellular function. Milo uses the large Western catalog of antibodies & can easily measure proteins irrespective of their location in or on a cell, making Milo the only platform with the versatility to detect diverse protein targets that are discovered in your RNA-sequencing runs.