Annexin V-FITC Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1 × 105 cells/mL were cultured. - Treated with various concentrations of M. integrifolia extract for 24, 48 and 72 h at 37 °C.	- After adding Annexin V–FITC and PI they were incubated for 15 min

Upstream tips
- 1 × 105 cells/mL were cultured. - Treated with various concentrations of M. integrifolia extract for 24, 48 and 72 h at 37 °C.
Protocol tips
- After adding Annexin V–FITC and PI they were incubated for 15 min

Publication protocol

Apoptotic cells were quantified with an Annexin V–FITC Apoptosis Detection Kit (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s protocol. Briefly, cells at 1 × 105 cells/mL were incubated with various concentrations (0, 30, 60 and 90 µg/mL) of M. integrifolia extract for 24, 48 and 72 h at 37 °C. Cells were then harvested and re-suspended in the binding buffer then stained with 2 µL Annexin V–FITC and 2 µL PI (propidium iodide) for 15 min at room temperature in the dark. The apoptotic index was immediately determined by flow cytometry.

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