Live / Dead assay mammalian cells - CHO-K1

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

Upstream tips
If the target cells are not completely lysed (as determined by microscopy), add another 5µl of Lysis Solution (10X).
Protocol tips
Add 50µl of CytoTox 96® Reagent to each well and cover the plate with foil or an opaque box to protect it from light and incubate for 30 minutes at room temperature
Downstream tips
If low % cytotoxicity is observed, increase the incubation time with the cytotoxic cells from 4 hours to 6–8 hours
Protocol tips
Add ethidium-calcein mixture, and incubate for 30 min

A shorter incubation time may be used if the dye concentrations or incubation temperature are increased.
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