Live / Dead assay mammalian cells - mouse keratinocytes

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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Found 2 matching solutions for this experiment

Live and Dead Cell Assay (Abcam)

Abcam

Upstream tips	Protocol tips	Downstream tips
	Add 4 μM of calcein acetomethoxy (calcein AM-5 μL) and 2 mM of ethidium-bromide. Incubate cells for 30-40 min at room temperature in the dark.	It is recommended to analyze the cells as quickly after staining as possible. The staining solution should not be exchanged for another buffer before data acquisition. Stained cells are stable for at least one hour at room temperature after staining. Live and Dead dye staining is lost if cells are fixed.

Protocol tips
Add 4 μM of calcein acetomethoxy (calcein AM-5 μL) and 2 mM of ethidium-bromide. Incubate cells for 30-40 min at room temperature in the dark.
Downstream tips
It is recommended to analyze the cells as quickly after staining as possible. The staining solution should not be exchanged for another buffer before data acquisition. Stained cells are stable for at least one hour at room temperature after staining. Live and Dead dye staining is lost if cells are fixed.

Manufacturer protocol Publication protocol 1 Relevant paper

LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	Add 1 μL of the reconstituted fluorescent reactive dye and incubate at room temperature or on ice for 30 minutes, protected from light.

Protocol tips
Add 1 μL of the reconstituted fluorescent reactive dye and incubate at room temperature or on ice for 30 minutes, protected from light.

Manufacturer protocol Publication protocol 1 Relevant paper

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