Cells were harvested and resuspended in PBS containing a 1:50 dilution of viability dye Zombie Aqua (30 (link)) for 10 minutes at room temperature and washed following the manufacturer’s protocol (BioLegend). Cells were then resuspended in 50 μL of fluorescence-activated cell sorting (FACS) buffer (BD Bioscience), and fluorophore-conjugated antibodies were added at 1:25 dilution, incubated for 15 minutes at 4°C. Antibodies were washed away twice by sequential centrifugation at 1400 RPM with FACS buffer and acquired in a BD Fortessa flow cytometer using FACSDiva software. Analysis of flow cytometry files was performed using FlowJo v 10.7 software.
Protocol detail
Flow Cytometry Viability Staining Protocol
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : UPMC Hillman Cancer Center
Other organizations : Essen University Hospital
Top 5 similar protocols
Variable analysis
independent variables
- Dilution of viability dye Zombie Aqua (1:50)
- Cell viability
- Cell harvesting and resuspension in PBS
- Incubation time (10 minutes)
- Incubation temperature (room temperature)
- Antibody dilution (1:25)
- Antibody incubation time (15 minutes)
- Antibody incubation temperature (4°C)
- FACS buffer used for cell resuspension and antibody washing
- Flow cytometry acquisition using BD Fortessa and FACSDiva software
- Data analysis using FlowJo v 10.7 software
- Manufacturer's protocol (BioLegend) for viability dye staining
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!