Biolegend
Zombie NIR Fixable Viability Kit
423105
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The aim was to discriminate between live and dead cells for further downstream analysis. Human CD4+ T cells isolated from PBMCs were used in the experiment.I chose this product as Biolegend offers quality products at good prices.
Flow cytometry
Human PBMCs
Stain cells (up to 5e5) in 50 ul PBS + Ab cocktail. Add Zombie UV at 1:500 dilution. Incubate for 15 min at RT or 30 min at 4deg. Wash with 150 ul FWB. Proceed to fixation and intracellular staining if needed.
Add Zombie UV dye to the PBS/Ab mix just before staining cells. Avoid any BSA/FCS in the staining medium.
I observed great separation of dead cells, which can be easily excluded from downstream analysis
NA
I do not recommend using this dye together with Alexa700 because of fluorescence spillover/compensation issues.
Good staining at high dilution
Need to use protein free medium. Not compatible with Alexa700
Good dye for live/dead cell discrimination.