Live / Dead assay mammalian cells - MCF-7 human breast cancer cells

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 the MTT Reagent to each well at a 1:10 ratio. For example, add 10 µL/well for a 96-well
plate or 25 µL/well for a 24-well plate.

Incubate the wells 2-4 hours or overnight at 37°C. Monitor the cells occasionally with an
inverted microscope for the presence of a purple precipitate
Protocol tips
Add Calcein AM (0.5 μM) and ethidium homodimer-1 (1 μM)

Incubate the cells for 30–45 minutes at room temperature
Protocol tips
Add 100 µl of assay solution to cells and incubate the mixture at 37 ºC for 15 min.
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