PE Annexin V Apoptosis Detection Kit I

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 5 µl of PE Annexin V and 5 µl 7-AAD. and incubate for 15 min at RT (25°C) in the dark.

Protocol tips
Add 5 µl of PE Annexin V and 5 µl 7-AAD. and incubate for 15 min at RT (25°C) in the dark.

Publication protocol

The relative cell size and DNA content were assessed by flow-cytometry analysis. At various time points, cells were harvested and fixed with 100% ethanol and then placed at –20 °C overnight. After fixation, cells were centrifuged and washed once with PBS containing 1% bovine serum albumin. For staining with DNA dye, cells were resuspended in 0.5 to 1 ml of PI solution containing RNase and incubated at 37 °C for 30 min, followed by overnight incubation at 4 °C. For Annexin V staining, 1 × 104 cells were collected and stained with Annexin V-FITC according to the manufacturer's instruction. The PI and Annexin V stained cells were collected and analyzed using BD flow-cytometry systems. The percentage of Annexin V-FITC positive cells was considered as the percentage of apoptotic cells.

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