Live / Dead assay mammalian cells - MC3T3

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
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 1.2 μL of 2 mM ethidium homodimer-1 (EthD-1) and 0.3 μL of 4 mM calcein AM.

Incubated for 30 min with 5% CO2 at 37 °C
Upstream tips
Cells were stained after 24 h of the culture
Protocol tips
Incubate cells for 10min at room temperature in the dark.
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