Annexin V-FITC Apoptosis Detection Kit

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- Cells were transfected and pre treated with CRH/Ucn2.	- After 24 h, the cells were used to assay apoptosis

Upstream tips
- Cells were transfected and pre treated with CRH/Ucn2.
Protocol tips
- After 24 h, the cells were used to assay apoptosis

Publication protocol

Annexin V–FITC/propidium iodide (PI) double staining was carried out according to a procedure described previously (Jin et al. 2011). Briefly, RM-1 cells were transfected with siRNA against cPLA2 and then treated with CRH/Ucn2 to detect apoptosis. In order to observe a significant antiapoptotic effect, TG (apoptosis inducer) was added to cells of the Ucn2 group to induce apoptosis in advance. After 24 h, the cells were harvested, washed, and resuspended in 0.5 ml PBS. Then cells were incubated for 15 min at room temperature in the dark in 10 μl Media Binding Reagent containing 1.25 μl Annexin V–FITC and 10 μl PI. Afterward, apoptosis was analyzed by fluorescence microscopy (Olympus) and flow cytometry (BD Biosciences, Franklin Lakes, NJ, USA).

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