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Annexin-V assay

WL Wenbin Li
YZ Yue Zhu
KZ Kelin Zhang
XY Xianhuan Yu
HL Haoming Lin
WW Wenrui Wu
YP Yaorong Peng
JS Jian Sun
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The ApopNexinTM FITC Apoptosis Detection Kit (Millipore) was used for the quantification of apoptotic cells in indicated cells, followed the manufacturer’s instruction. Briefly, indicated treated cells were firstly washed with PBS twice and then responded with the Annexin-V binding solution, subsequently added 150 μl of the Annexin-V antibody in Binding Buffer and incubated for 15 min. Afterward, addition of 1.5 μl of PI at the concentration as 1 mg/ml and a further incubation for 5 min were processed. After washing with the Annexin-V Binding Buffer, positive Annexin-V staining was visualized under a Flow Cytometer equipped with two panels for fluorescein isothiocyanate (excitation: 490 nm, emission: 525 nm), and PI staining was assessed with the filter for Texas red (excitation: 570 nm, emission: 610 nm).

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