Annexin V-FITC Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 5 µl of PE Annexin V and 5 µl 7-AAD and incubate for 15 minutes in dark

Protocol tips
Add 5 µl of PE Annexin V and 5 µl 7-AAD and incubate for 15 minutes in dark

Publication protocol

After culture in serum-free media for 24 h, cells were harvested and washed with cold PBS. Levels of phosphatidylserine on the cell surface were estimated by using the Annexin V-FITC and PI apoptosis detection kit according to the manufacturer's instructions (Beyotime). Apoptotic cells were analyzed in a flow cytometer (Beckman Coulter, Brea, CA, USA) using FL1 (excitation 488 nm, green) and FL3 (excitation 585 nm, red) channels. A minimum of 10,000 events were collected for each sample. Results are expressed as the percentage of apoptotic cells relative to total cells.

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