Consistent Fixable Viability Dye From BioLegend

August 11, 2023

George Washington University

Overall

Quality of Results

Ease-of-Optimization

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Company:

BioLegend

Product Name:

Zombie Green™ Fixable Viability Kit

Catalog Number:

423112

Clone Number:

N/A

My research involves significant flow staining of tumor cell lines after exposing them to treatments that induce cell killing.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Human dendritic cells

Primary Incubation

0.1uL Zombie Green viability stain in 100uL PBS, incubated at room temp for 15 mins.

Blocking Agent

Human TruStain Fc block, 5uL diluted in 95uL FACS buffer (5% FBS in DPBS), incubated at 4C for 10 mins.

Secondary Incubation

Desired antibody (as needed).

Tertiary Incubation

Cytofix buffer, 100uL for 20 mins at 4 degrees C.

Detection

Flow cytometry in a beckman coulter cytoflex cytometer.

Results Summary

This product helped me identify live populations, and has helped me consistently identify live populations with and without fixing cells. I consistently see distinct live and dead populations, and sometimes a third "half-dead" population (not pictured) if a significant portion of cells are marked for apoptotic clearance.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Great fixable viability dye, only need 0.1uL/sample.

The Bad

Huge overlap with PE, must build panels with caution.

The Bottom Line

It's a great fixable viability dye if you don't have anything in your panel with too much overlap. BioLegend also makes other Zombie dyes, which work with just as much confidence in case you do!

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