FITC Annexin V Apoptosis Detection Kit with 7-AAD

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were pre treated with 8 μM sorafenib for 24h. - 1 × 10^6 cells were used for the assay.	- Add FITC Annexin V and 7-AAD Viability Staining Solution and incubate for 15 min at RT.

Upstream tips
- Cells were pre treated with 8 μM sorafenib for 24h. - 1 × 10^6 cells were used for the assay.
Protocol tips
- Add FITC Annexin V and 7-AAD Viability Staining Solution and incubate for 15 min at RT.

Publication protocol

Many agents, including the clinically useful sorafenib, can induce apoptosis [20]. Following 24-h treatment with 8 μM sorafenib, PANC-1–derived clones were trypsinized, washed with cold PBS, and resuspended in PBS. The advantage of 7-amino-actinomycin D (7-AAD) over propidium iodide is that there is minimal spectral overlap between the emissions. We used a fluorescein isothiocyanate (FITC) Annexin V Apoptosis Detection Kit with 7-AAD (#640922; BioLegend, USA) according to the manufacturer’s instructions. Briefly, 5 μL FITC–annexin V and 5 μL 7-AAD Viability Staining Solution (#00-6993; eBioscience, USA) were added to a 100-μL cell suspension (1 × 106 cells) in binding buffer. The cells were gently vortexed and incubated for 15 min at room temperature in the dark, and 400 μL binding buffer was added for flow cytometric analysis using a FACScan Flow Cytometer (Becton Dickinson).

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Manufacturer protocol

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