Annexin V-FITC Apoptosis Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells are treated with 50 nM of oleandrin for 0, 24 and 48 h.	- Always stain cells with Annexin before fixation (makes cell membrane leaky)

Upstream tips
- Cells are treated with 50 nM of oleandrin for 0, 24 and 48 h.
Protocol tips
- Always stain cells with Annexin before fixation (makes cell membrane leaky)

Publication protocol

U2OS and SaOS-2 cells were seeded into 6-well plates and adhered overnight. When a confluence of 70–80 % was reached, both cell lines were exposed to 50 nM of oleandrin for 0, 24 and 48 h. Subsequently, the cells were collected and re-suspended in 500 μl of 1× binding buffer. Five microliters of annexin V-FITC and 5 μl of propidium iodide (PI) were added to all of the samples, which were then incubated at room temperature for 5 min in the dark according to the manufacturer’s protocol from the Annexin V-FITC apoptosis detection kit (BioVision Co, California, USA). The fluorescence intensity of the cells was immediately analyzed by flow cytometry.

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Manufacturer protocol

Download the product protocol from Biovision for Annexin V-FITC Apoptosis Kit below.

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