Zombie Aqua™ Fixable Viability Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Wash cells with PBS buffer (no Tris buffer and protein free). Dilute Zombie Aqua™ dye at 1:100-1000 in PBS. Resuspend 1-10 x 10 cells in diluted 100 µl Zombie Aqua™ solution. Incubate the cells at room temperature, in the dark, for 15-30 minutes.

Protocol tips
Wash cells with PBS buffer (no Tris buffer and protein free). Dilute Zombie Aqua™ dye at 1:100-1000 in PBS. Resuspend 1-10 x 10 cells in diluted 100 µl Zombie Aqua™ solution. Incubate the cells at room temperature, in the dark, for 15-30 minutes.

Publication protocol

PBMCs (1 × 106 cells per group) were stimulated with S. aureus isolates under the conditions indicated in the relevant figure legends. For intracellular cytokine staining, 3 μg/ml of brefeldin A (eBioscience) was added after 6 h of stimulation, and the stimulation was continued for an additional 12 h. Dead cells were excluded from the analysis using a Zombie Aqua fixable viability kit (BioLegend). Cells were washed in PBS containing 2% FBS and 2 mM EDTA, blocked with 10% normal human serum, and stained for CD3, CD14, and CD19. Cells were fixed and permeabilized using the Cytofix/Cytoperm kit (BD Biosciences) and stained for IL-10 and TNF-α. For phospho-flow, PBMCs were stimulated for 30 min, fixed with Fix Buffer I (BD Biosciences), stained for extracellular markers, permeabilized using Perm/Wash Buffer I (BD Biosciences), and stained for the intracellular molecules of interest. Events were acquired on a LSRII Fortessa (BD), and doublets were excluded based on forward side scatter-area (FSC-A)/forward side scatter-height (FSC-H). Data analysis was performed using FlowJo version 10.x (Tree Star Inc.).

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 bacteria - Staphylococcus aureus 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 bacteria - Staphylococcus aureus 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.