Annexin V-FITC Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 5 µl of FITC Annexin V and 5 µl PI and incubate for 15 min at room temperature in the dark.

Protocol tips
Add 5 µl of FITC Annexin V and 5 µl PI and incubate for 15 min at room temperature in the dark.

Publication protocol

MG-63 and U2OS cells seeded in 12-well plates (1.5 × 105 cells/well) were treated with increasing concentrations of curcumin for 24 h before harvest. Cells were assessed for apoptosis using an Apoptosis Detection kit (eBioscience, San Diego, CA, USA) according to manufacturer’s protocol. Briefly, cells suspended in binding buffer were incubated with Annexin V-FITC and propidium iodide (Becton Dickinson, Mountain View, CA, USA) for 15 min at room temperature in the dark. Cells were then analyzed by flow cytometry performed on a FACScalibur cell analyser using CellQuest software (Becton Dickinson, Mountain View, CA, USA). Cells undergoing apoptosis were defined as those positive for Annexin V.

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Manufacturer protocol

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