Live / Dead assay mammalian cells - H9C2

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 3 matching solutions for this experiment

Protocol tips
Add test compounds and equilibrate the plate and its contents at room temperature for approximately 15 minutes.

Add CellTiter-Glo® Reagent and mix contents for 2 minutes on an orbital shaker to induce cell lysis.

Allow the plate to incubate at room temperature for 10 minutes to stabilize luminescent signal
Protocol tips
Add 100 µl of the Calcein-AM/EthD-III to each well.

Incubate the samples at room temperature for 30–45 minutes
Protocol tips
Add ethidium-calcein mixture, and incubate for 30 min in a 37 °C incubator
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