FITC Annexin V Apoptosis Detection Kit I

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1×10^5 cells/ml cells were seeded for an assay. - BAK cells were added to HeLa cells after 6h and incubated for 20h.	- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark

Upstream tips
- 1×10^5 cells/ml cells were seeded for an assay. - BAK cells were added to HeLa cells after 6h and incubated for 20h.
Protocol tips
- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark

Publication protocol

HeLa cells (1×105 cells/ml) were incubated in 12-well plates with a volume of 1 ml in each well. After 6 h, when the cells were adherent, BAK cells (2×106 cells/ml, 1 ml) were added to the adherent HeLa cells (2 ml total per well) and incubated for 20 h. Unstimulated PBMCs were added to HeLa cells at the same E/T ratio to serve as the negative control. The wells containing HeLa cells without effector cells were supplemented with 1 ml RPMI-1640 medium to serve as the blank control. Following incubation, the suspended cells (BAK and NK cells) were washed three times with PBS to remove all effector cells. The HeLa cells were trypsinized and washed with PBS, then collected and stained using the FITC Annexin V Apoptosis Detection kit I (BD Biosciences, San Diego, CA, USA) according to the manufacturer’s instructions. The stained HeLa cells were analyzed by fluorescent-activated cell sorting (FACS) using a BD LSR II Flow Cytometer (BD Biosciences).

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Download the product protocol from BD Biosciences for FITC Annexin V Apoptosis Detection Kit I below.

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