Zombie Aqua™ Fixable Viability Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Dilute Zombie Aqua™ dye at 1:100-1000 in PBS. Incubate the cells at room temperature, in the dark, for 15-30 minutes

Protocol tips
Dilute Zombie Aqua™ dye at 1:100-1000 in PBS. Incubate the cells at room temperature, in the dark, for 15-30 minutes

Publication protocol

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   Login or join for free to view the full paper.

Reviews

Discussion

Papers

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.

Manufacturer protocol

Download the product protocol from BioLegend for Zombie Aqua™ Fixable Viability Kit below.

Download manufacturer protocol

Videos

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.