Apoptosis assay cell type - OV2008

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample 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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Protocol tips
OVCAR-3, A2780/CP70 and IOSE-364 cells were seeded in 24-well plates at 1 × 105 cells/well and incubated overnight. Cells were treated with various concentrations (0–4 μM) of ChK for 24 h. After treatment, cells were stained with 10 μg/mL Hoechst 33342 (Sigma, St. Louis, MO) in PBS for 10 min in the dark at 37 °C. Cell apoptosis was examined under a fluorescence microscope (ZEISS), and data were collected from three independent experiments.
Protocol tips
Add 5 µl of FITC Annexin V and 5 µl PI and incubate for 15min at RT.
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