Live / Dead assay mammalian cells - rat nucleus pulposus

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

Protocol tips
Add 10 μl CCK-8 solution incubate for 2 h.

Downstream tips
Absorbance of each wells was measured at 450 nm

To measure the absorbance later, add 10 μl of 1% w/v SDS
or 0.1 M HCl to each well, cover the plate and store it with
protection from light at room temperature. No absorbance
change should be observed for 24 hours
Upstream tips
Wash cells 2–3 times with PBS
Protocol tips
Add calcein AM: 2 μM; EthD-1: 4 μM

Incubate the cells for 40 minutes at room temperature
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