FITC Annexin V Apoptosis Detection Kit I

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 1 × 10^6 cells/ml. were used for assay.	- Annexin V and propidium iodide (PI) were added and incubated in dark for 15 minutes - Cells were fixed, washed and treated with RNase and incubated for 15 minutes at 37°C.

Upstream tips
- 1 × 10^6 cells/ml. were used for assay.
Protocol tips
- Annexin V and propidium iodide (PI) were added and incubated in dark for 15 minutes - Cells were fixed, washed and treated with RNase and incubated for 15 minutes at 37°C.

Publication protocol

The procedure was performed with Annexin V-FITC Apoptosis Detection Kit (BD Pharmingen, USA) using a modified protocol by Rieger et al. [27]. Briefly, upon completion of treatments, cells were harvested and washed with binding buffer. Cells were counted to obtain a final concentration of 1 × 106 cells/ml. Then Annexin V and propidium iodide (PI) were added and incubated in dark for 15 minutes. After washing, cell suspension was fixed with 1% formaldehyde for 10 minutes on ice. Subsequently, washing was performed twice with binding buffer followed by addition of RNase (EMD Biosciences, USA) which then incubated for 15 minutes at 37°C. Finally, samples were washed and ready for analysis with FACSCalibur flow cytometer (BD Biosciences, USA) and the software Cell Quest Pro.

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