Cell apoptosis was detected using flow cytometry with an Annexin V-phycoerythrin (PE)/7-aminoactinomycin D (7AAD) kit (BD Biosciences, NJ, USA) as published in our paper.24 (link) The cells and culture media were collected and centrifuged, and the supernatant was discarded. Next, the cells were rinsed with PBS 3 times. The cells were resuspended in 10 mmol/L binding buffer and incubated with Annexin V-PE/7AAD in the dark for 15 min. Groups of 10,000 cells were analysed using a BD LSRFortessa (BD Biosciences).
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Annexin V-PE/7AAD Flow Cytometry for Apoptosis
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Corresponding Organization : Shandong University
Other organizations : Shandong University of Traditional Chinese Medicine
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Variable analysis
independent variables
- Cell treatment (exposure to experimental conditions)
- Cell apoptosis measured using flow cytometry with Annexin V-PE/7AAD staining
- Cell culture media
- PBS for cell rinsing
- Binding buffer concentration (10 mmol/L)
- Incubation time with Annexin V-PE/7AAD (15 min)
- Number of cells analyzed (10,000)
- Positive control: Cells exposed to known apoptosis-inducing conditions
- Negative control: Untreated cells
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