PE Annexin V Apoptosis Detection Kit I

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- 5×10^5 cells were seeded. - Pretreated with 0.3 µM imatinib for 24 h	- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark

Upstream tips
- 5×10^5 cells were seeded. - Pretreated with 0.3 µM imatinib for 24 h
Protocol tips
- Treated with FITC Annexin V and PI and incubated for 15 min at RT (25°C) in the dark

Publication protocol

For the apoptosis assay, 5×105 K562-miR-181a and K562-EGFP cells were exposed to 0.3 µM imatinib for 24 h, and subsequently, cell apoptosis was detected with an apoptosis kit (BD Biosciences). The cells were collected, washed and resuspended in binding buffer at a density of 1×106 cells/ml. Next, 1×105 cells were incubated for 15 min with 5 µl phycoerythrin (PE)-Annexin V and 5 µl 7-aminoactinomycin D at room temperature in the dark, and analyzed using a FACSCanto II flow cytometer (BD Biosciences) 1 h later.

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