Flow cytometry to determine the apoptosis rate in H9c2 cells
This protocol is extracted from research article:
Os Efeitos da Doxorrubicina na Biossíntese e no Metabolismo do Heme em Cardiomiócitos
Arq Bras Cardiol, Feb 19, 2021; DOI: 10.36660/abc.20190437

Apoptosis was assessed using the Annexin V-FITC/PI Apoptosis Detection kit (KeyGEN, China) according to the manufacturer’s protocol. After the intervention, approximately 1×105 H9c2 cells of each group were washed and digested with trypsin, and then resuspended with 1x PBS (4°C) and centrifuged at 2,000 rpm for 5-10 minutes to wash the cells. The cells were resuspended in 500μl of the buffer, followed by the addition of 5 μl of Annexin V-FITC and 5 μl of PI. The cells were incubated in the dark for 15 minutes at room temperature. Next, the cells of each group were examined for apoptosis rate by flow cytometry (BD Accuri™ C6). The experiment was repeated three times. The apoptotic rate was quantified as the percentage of cells stained with Annexin V.

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