Product: Zombie Green™ Fixable Viability Kit

Catalog No.: 423112

Lucy LeBlanc, Graduate Research Assistant University of Texas, Austin

Excellent Alternative to Classical Viability Dyes (Zombie Green™ Fixable Viability Kit)
Overall:	Product Quality:	Ease of Use:
Gives distinct non-viable vs. viable peaks, easy to integrate into existing flow protocols as its excitation/emission spectra are pretty sharp and similar to FITC; compatible with sequential antibody or annexin V staining without the need for washing cells in between.

Experimental Design

Application:	Flow Cytometry
Cells used:	Murine embryonic stem cells
Brief Protocol:	Used accutase to detach the monolayer and generate a single cell suspension. Washed with protein-free PBS after centrifugation. Resuspended in annexin V binding buffer (actually, any buffer works here as long as it doesn't contain BSA or something). Generated positive control by treating cells with 0.1% Triton. Pre-diluted Zombie Green 1:10 in PBS and then treated cell suspension 1:100 with the pre-dilution, so ultimately a 1:1000 dilution of ZG was sufficient (therefore, the kit is quite economical). Incubated in the dark, then added annexin V, incubated another 15 mins, centrifuged and resuspended in buffer followed by flow cytometry.
Results Summary:	Gating revealed distinct early apoptotic vs. necrotic populations. Very little background/noise (early apoptotic sample showed fluorescence almost equivalent to the unstained control, for example, as expected).
Additional Notes:	Keep everything protected in foil and work in low light, since this is a fluorophore, as usual.

Data Image Click on the image for full size data.

Publication Status This data has not been published.

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