The cells were trypsinized and fixed in 75% ethanol at 4°C for 25–30 min. The cell pellets were subsequently harvested and incubated using an Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit (Invitrogen; Thermo Fisher Scientific, Inc.). Cellular apoptosis was evaluated via flow cytometry (Phoenix Flow Systems, San Diego, CA, USA) within 5 min and analyzed using a flow cytometer (Coulter EPICS XL-MCL FACScan, BD Biosciences, Franklin Lakes, NJ, USA). The data was analyzed using the MultiCycle Software for Windows version 5.0 (Phoenix Flow Systems, San Diego, CA, USA).
Annexin 5 fluorescein isothiocyanate propidium iodide apoptosis detection kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit is a laboratory reagent used to detect and quantify apoptosis in cells. It contains Annexin V, a protein that binds to phosphatidylserine, and propidium iodide, a DNA-binding dye. Together, these components can identify different stages of apoptosis in a cell population.
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2 protocols using annexin 5 fluorescein isothiocyanate propidium iodide apoptosis detection kit
1
Evaluating Cellular Apoptosis by Flow Cytometry
2
Quantifying Early and Late Apoptosis in ARPE19 Cells
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The early and late apoptosis rates in ARPE19 with different treatments were determined using an Annexin V-Fluorescein Isothiocyanate/Propidium Iodide Apoptosis Detection kit according to the the instructions of manufacturers (Invitrogen, USA). Briefly, the cells were resuspended in the binding buffer to a concentration of 10 6 cells/ml. Then, 5 µL Annexin V-fluorescein isothiocyanate and propidium iodide were added, followed by incubation at room temperature for 15 min in the dark. Ultimately, the cell fluorescence was detected on FACScan by flow cytometry within 1 h. FlowJo software was used to analyze the flow data.
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