Cell cytotoxicity / Proliferation assay cell type - OVCAR-3

Cell cytotoxicity assays measure the ability of certain compounds or chemical mediators to reduce the viability of the cells. The term cell cytotoxicity assay can sometimes be used interchangeably with cell proliferation assay. Healthy living cells can be identified by the use of formazan dyes, protease biomarkers or by measuring ATP content. The formazan dyes are chromogenic products formed by the reduction of tetrazolium salts by dehydrogenases, such as lactate dehydrogenase (LDH) and reductases that are released during cell death. Common tetrazolium salts include INT, MTT, MTS and XTT. Cell cytotoxicity can also be measured by using the SRB and WST-1 assays. These assays can usually be used in a high-throughput fashion and can be quantitated by measuring absorbance, colorimetry or luminescence. All these assays require similar numbers of cell plating at the initiation, a time course of treatment with the cytotoxic agent and at least triplicates for each condition at every point of analysis. Cell shrinkage, plasma membrane blebbing, cell detachment, externalization of phosphatidylserine, nuclear condensation and ultimately DNA fragmentation are well-described features of apoptosis. The assays that rely on cell membrane integrity for their function, may not be able to quantify early apoptosis. Therefore, in order to distinguish early apoptotic vs. late apoptotic or necrotic cells, additional flow cytometry techniques can be used. A combination of Annexin V and PI (propidium iodide) can be used to distinguish early (Annexin V+/PI-) and late apoptotic (Annexin V+/PI+) cells. Sometimes, caspase assays are used in order to differentiate the stages of apoptosis.

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Found 3 matching solutions for this experiment

BrdU Cell Proliferation Assay Kit

Cell Signaling Technology

Upstream tips
-After cells were attached, they are treated without or with thyrostimulin and/or effector inhibitors for 24 h.
Protocol tips
- Incubation time with BrdU was for 4h.

- After addition of AlamarBlue, cells were incubated for 3 h at 37 °C.
Protocol tips
- Plates are incubated for an appropriate length of time (e.g. 0, 24, 48, or 72 hours) in the incubator.

- Cells were incubated for 1.5 h at 37 °C after addition of CCK-8 reagent
Upstream tips
- 1000 cells/ well were platted in a 96 well plate
Protocol tips
- XTT reagent was added to each well, every 24 h until 96 h and incubated for 5 h.
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