Annexin V-FITC Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Prepare 1x Binding Buffer by diluting 1 ml of the 10x Binding Buffer with 9 ml of deionized water. - Dissolve the staurosporine in DMSO to a concentration of 100 mg/ml	- 1 × 10^4 cells were collected and stained with Annexin V-FITC

Upstream tips
- Prepare 1x Binding Buffer by diluting 1 ml of the 10x Binding Buffer with 9 ml of deionized water. - Dissolve the staurosporine in DMSO to a concentration of 100 mg/ml
Protocol tips
- 1 × 10^4 cells were collected and stained with Annexin V-FITC

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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