eBioscience™ Annexin V-FITC Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1.3 × 10^5 cells were seeded and incubated with DCM-DS (12.5-50 μg/mL) for 24 h	- Annexin V-FITC is treated on cells and incubated for 10 min at room temperature in dark

Upstream tips
- 1.3 × 10^5 cells were seeded and incubated with DCM-DS (12.5-50 μg/mL) for 24 h
Protocol tips
- Annexin V-FITC is treated on cells and incubated for 10 min at room temperature in dark

Publication protocol

Experiment was carried out according to the manufacturer’s instructions (Annexin V-FITC kit, eBioscience, Vienna, Austria) with slight modification. Cells were seeded in 6-well plates at 1.3 × 105 cells per well in 3 mL of complete growth culture media, incubated for 24 hours and treated with DCM-DS (12.5-50 μg/mL). Following incubation, the floating cells were collected and the adherent cells were trypsinized to detach the cells. Cells were counted and a volume of media containing 1 × 105 cells (both viable and death cells) was centrifuged at 100 × g to obtain a pellet. Next, 195 μL of 1× assay buffer and 5 μL of Annexin V-fluorescein isothiocyanate (FITC) were added to the pellet. The samples were mixed by gentle tapping. Following 10 min incubation at room temperature in the dark, 300 μL of 1× assay buffer and 10 μL of PI (20 μg/mL) were added, and the samples were analysed immediately by FACSCalibur flow cytometer (Becton Dickinson, CA, USA). For each sample, 10000 events were collected. The results were analysed by using FlowJo software.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for eBioscience™ Annexin V-FITC Apoptosis Detection Kit below.

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