FITC Annexin V Apoptosis Detection Kit I

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 2 × 10^5 cells were cultured	- Annexin V‐FITC is treated for 30 min and then with propidium iodide (PI) for 5 min at room temperature,

Upstream tips
- 2 × 10^5 cells were cultured
Protocol tips
- Annexin V‐FITC is treated for 30 min and then with propidium iodide (PI) for 5 min at room temperature,

Publication protocol

Apoptosis induced by TRAIL and 3‐MA was detected with an Annexin V‐FITC apoptosis detection kit according to the manufacturer's instructions (BD Biosciences Pharmingen, CA). Briefly, 2 × 105 cells were washed once with ice cold phosphate‐buffered saline (PBS, pH 7.4) and re‐suspended in 200 μl binding buffer. Cells were incubated with Annexin V‐FITC for 30 min and then with propidium iodide (PI) for 5 min at room temperature, and 400 μl binding buffer was added before testing. Apoptotic cells were detected by flow cytometry (FACS Calibur, Becton Dickinson). Cell viability was quantified by a 3‐(4, 5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxy‐ methoxyphenyl)‐2‐(4‐sulfophenyl)‐2H‐tetrazolium (MTT) assay (Sigma, St. Louis, MO).

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