Reliable Zombie NIR™ Fixable Viability Kit From BioLegend

Department of Immunology and Microbiology
Scripps Research
Graduate Student

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Company:

BioLegend

Product Name:

Zombie NIR™ Fixable Viability Kit

Catalog Number:

423106

Clone Number:

RUO

The Zombie NIR viability kit works well for identifying dead cells from cell suspensions isolated from mouse lungs, spleen, and lymph nodes. I recommend storing at -20C to ensure the dye works well over multiple freeze-thaw cycles. Do not store at 4C. Despite being excited by the red laser, this dye has worked for me even with APC, APC-fire750, and APC-810 in the same panel.

Experimental Design and Results Summary

Applications

Flow cytometry

Sample

Mouse lung, mediastinal lymphnode, spleens

Primary Incubation

1:1000, 20 minutes at 4 degrees C in the dark

Blocking Agent

Fc Shield

Secondary Incubation

BD cytofix/cytoperm; 100 ul for 20 mins at 4 degrees C

Tertiary Incubation

None

Detection

5 laser Aurora Flow cytometer

Results Summary

I used this product to identify Live (Zombie NIR negative population) immune cells from cells isolated from the lung, spleen and lymph nodes of mice. Cells were isolated from BALF using 1 ml of 1% BSA in PBS and from lungs through collagenase digestion; RBC lysis was performed using ACK buffer for all samples. Cells were then stained with viability dye in Fc block followed by extracellular antibodies such as CD45, TCR beta, CD4, CD8, and so on. After fixing and permeabilizing, I have also stained for intracellular cytokine expression. I have been impressed by the ability of the aurora to distinguish this dye’s signal (at R6) from that of APC-fire 750 (R7) and APC-fire 810 (R8).

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Cheap and good signal.

The Bad

Must be stored at -20 degrees C; takes time to thaw.

The Bottom Line

I would highly recommend this live-dead stain since it uses a channel that is highly compatible with most other common fluorophores.

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