|Dilute Zombie Aqua™ dye at 1:100-1000 in PBS.
Incubate the cells at room temperature, in the dark, for 15-30 minutes
Cells were thawed, washed 2x in pre-warmed cell culture medium and rested for 2h at 37°C, 5% CO2. Cells were washed in pure PBS and stained with zombie aqua (BioLegend), washed 1x in pure PBS and stained with CD24 and CD38 antibodies (Suppl. Table 4) for 15min at 4°C. After washing in pre-warmed cell culture medium cells were re-suspended in 250μl warm cell culture medium and immediately stimulated as described in the cell stimulation section. Stimulation was stopped by adding 150μl of 4% PFA and incubated for 15min at RT. Cells were washed with pure PBS and permeabilized in methanol at −80°C overnight. After 2x washing in pure PBS, cells were stained with an intracellular staining cocktail of antibodies specific for phosphorylated signaling molecules and additional phenotyping markers (Suppl. Table 4), washed, and finally stained with AF488-labeled goat anti-rabbit antibody (Suppl. Table 4) before analysis on an LSRII flow cytometer (BD Biosciences). Full paper
or join for free
to view the full paper.
Zombie Aqua™ Fixable Viability Kit from BioLegend has not yet been reviewed for this experiment
We'd love it if you would be the first to write a review!
Start your discussion
Share your thoughts or question with experts in your field
Start a discussion
Check out relevant papers found by Labettor's AI that are relevant for performing Live / Dead assay mammalian cells - human peripheral blood mononuclear cells using Zombie Aqua™ Fixable Viability Kit from BioLegend.
Mass cytometry analysis shows that a novel memory phenotype B cell is expanded in multiple myeloma
Check out videos that might be relevant for performing Live / Dead assay mammalian cells - human peripheral blood mononuclear cells using Zombie Aqua™ Fixable Viability Kit from BioLegend. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.
We haven't found any additional videos for this experiment / product combination yet.