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).
Annexin 5 phycoerythrin pe 7 aminoactinomycin d 7aad kit
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The Annexin V-phycoerythrin (PE)/7-aminoactinomycin D (7AAD) kit is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. The kit contains Annexin V-PE, which binds to phosphatidylserine, and 7AAD, a DNA-binding dye. Together, these two components allow for the identification and differentiation of viable, early apoptotic, and late apoptotic/necrotic cells through flow cytometry analysis.
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3 protocols using annexin 5 phycoerythrin pe 7 aminoactinomycin d 7aad kit
1
Annexin V-PE/7AAD Flow Cytometry for Apoptosis
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Apoptosis Analysis of Glioma Cells
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An Annexin V‐phycoerythrin (PE)/7‐aminoactinomycin D (7‐AAD) kit (BD Biosciences) was used to examine cell apoptosis. The 7‐AAD is a standard flow cytometry probe used to distinguish living and non‐living cells. In short, the glioma cells were resuspended in 1 × binding buffer and incubated with Annexin V‐PE/7‐AAD in the dark at 25℃ for 15 min. Apoptosis of cells was then analyzed using a FACS Canto™II flow cytometer (BD Biosciences) within 1 h.
3
Quantifying Apoptosis by Flow Cytometry
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Cell apoptosis was measured using flow cytometry using Annexin V-phycoerythrin (PE)/7-aminoactinomycin d (7AAD) kit (BD Biosciences, NJ, USA) following the manufacturer’s protocol. In brief, cells in culture media were collected by centrifugation, and then the cell pellets were washed with 1× phosphate-buffered saline three times. The collected cells were then stained with annexin V-PE/7AAD for 15 min in the dark at room temperature according to the manufacturer’s instructions. Groups of 10,000 cells were analyzed using a BD LSRFortessaTM (BD Biosciences, NJ, USA).
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