Dead Cell Apoptosis Kit with Annexin V FITC and PI

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 2 × 10^4 cells/well were seededfor assay. - cells were treated with LPLI	- Cells are stained with PI and annexin V-FITC and incubated on ice for 10–15 min

Upstream tips
- 2 × 10^4 cells/well were seededfor assay. - cells were treated with LPLI
Protocol tips
- Cells are stained with PI and annexin V-FITC and incubated on ice for 10–15 min

Publication protocol

For the apoptosis assay, the detached oral cancer cells treated with LPLI were washed with PBS and stained with propidium iodide (PI) and annexin V-FITC (Invitrogen, V13241) on ice for 10–15 min. The stained cells were analyzed using a FACScan flow cytometer (Becton Dickinson) and the number of apoptotic cells was quantified using FlowJo software (Tree Star). Alternatively, cells were seeded into 96-well white plate (2 × 104 cells/well) for overnight and exposed to LPLI in the presence or absence of autophagy inhibitors. The cells were then recovered for 24 h and lyzed with Caspase-Glo 3/7 (Promega, G8091) to measure luminescent signal for 1 h. The net relative luminescence units (RLU) between time 0 and 1h were used to reflect caspase 3/7 activity in treated cells.

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