Annexin V-FITC Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Add 5 ml of Annexin V FITC Conjugate and 10 ml of Propidium Iodide Solution to each cell suspension and incubate the tubes at room temperature for exactly 10 minutes and protect from light.

Protocol tips
Add 5 ml of Annexin V FITC Conjugate and 10 ml of Propidium Iodide Solution to each cell suspension and incubate the tubes at room temperature for exactly 10 minutes and protect from light.

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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