Cell-APOPercentage™ Apoptosis Assay

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 3 × 10^4 were seeded in 96 well plate.	- Cells were cultured in culture medium with and without fresh medium containing 5% FBS (as control) or 5% ascite in presence or not of H2O2 for 6 hours before adding APOPercentage Dye.	- Wells were shaken for 20 minutes before being read at 540 nm

Upstream tips
- 3 × 10^4 were seeded in 96 well plate.
Protocol tips
- Cells were cultured in culture medium with and without fresh medium containing 5% FBS (as control) or 5% ascite in presence or not of H2O2 for 6 hours before adding APOPercentage Dye.
Downstream tips
- Wells were shaken for 20 minutes before being read at 540 nm

Publication protocol

Apoptosis was measured with the apoptosis assay APOPercentage from Biocolor as previously described [27]. Briefly, 3 × 104 SKOV3 cells were seeded in gelatine-coated 96 wells and incubated in 100 μl of medium at 37°C, 5% CO2 until confluence was reached (24h). The incubation culture medium was then removed and replaced by 100 μl of fresh medium containing 5% FBS (as control) or 5% ascite in presence or not of H2O2 for 6 hours before addition of 5 μl of APOPercentage Dye for 30 minutes at 37°C and 5% CO2. Culture medium was then removed, and cells were gently washed with PBS to remove unbound dye. Then, either red cells were counted under microscopy or 100 μl of Dye releasing reagent was added to the wells to extract bound dye. Wells were shaken for 20 minutes before being read at 540 nm.

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Manufacturer protocol

Download the product protocol from Biocolor for Cell-APOPercentage™ Apoptosis Assay below.

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