After treatment with or without MO-B (10, 20 and 40 µM) for 24 h, cells were exposed to 1,000 µM H2O2 for 6 h and then washed with ice-cold PBS. Apoptosis was analyzed using an Annexin V-fluorescein isothiocyanate/propidium iodide Apoptosis Detection Kit (Beyotime Institute of Biotechnology). Cells were stained according to the manufacturer's instructions. The proportion of apoptotic cells in 1×105 labeled cells was quantified using a BD Accuri C6 Plus flow cytometry and BD Accuri C6 software (BD Biosciences).
Annexin 5 fluorescein isothiocyanate propidium iodide apoptosis detection kit
Manufactured by Beyotime
Sourced in China
The Annexin V-fluorescein isothiocyanate/propidium iodide Apoptosis Detection Kit is a laboratory instrument used to detect and quantify apoptosis (programmed cell death) in cell samples. The kit utilizes fluorescently-labeled Annexin V and propidium iodide dyes to differentiate between viable, apoptotic, and necrotic cells.
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8 protocols using annexin 5 fluorescein isothiocyanate propidium iodide apoptosis detection kit
1
Oxidative Stress-Induced Apoptosis Assay
2
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis was determined by Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit (Beyotime, Shanghai, china). Trypsin-digested cells in the absence of ethylenediaminetetraacetic acid were washed with PBS, resuspended in 100 µL of 1 × binding buffer, and then added with 5 μL of Annexin V-FITC and 10 μL of PI. After 15 min incubation in the dark, apoptosis was detected with a flow cytometer (BD Biosciences, Franklin Lake, Franklin Lake, New Jersey, USA), and data analysis was performed using FlowJo software (Stanford University, San Francisco, California, USA).
3
Apoptosis Assay of Glioma Cells
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An annexin V–fluorescein isothiocyanate/propidium iodide apoptosis detection kit (Beyotime BioTech) was used to measure apoptosis of glioma cells, as described by the manufacturer. The glioma cells were maintained in plates and then transfected with miR-1272 mimics or inhibitors and their negative controls. After incubation for 24 h, the harvested glioma cells were stained with Annexin V-FITC and PI and were then analyzed by flow cytometry using WinMDI 2.9 software (The Scripps Research Institute, La Jolla, CA, USA).
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Annexin V-FITC/PI Apoptosis Assay
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After transfection for 48 h, apoptotic dMSCs were trypsinized and stained using an Annexin V-fluorescein isothiocyanate/propidium iodide apoptosis detection kit (C1062M; Beyotime, Shanghai, China), according to the manufacturer’s instructions. A flow cytometer (BD Biosciences, USA) was used to calculate cell apoptosis, and the results were analyzed using CellQuest software.
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Apoptosis Analysis of Y79 and WERI-Rb-1 Cells
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The apoptosis of Y79 and WERI-Rb-1 cells was analyzed as per the instructions of an Annexin V-fluorescein isothiocyanate/propidium iodide Apoptosis Detection Kit (Beyotime) and estimated by FACScan® flow cytometry (BD Biosciences, San Jose, CA, USA).
6
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis was determined using the annexin V-fluorescein isothiocyanate/propidium iodide (PI) apoptosis detection kit (Beyotime Biotechnology) according to the manufacturer’s instructions. Briefly, the cells were cultured in chamber slides (3×104 cells per well), allowed to attach overnight and exposed to various experimental conditions. After treatment for 72 h, the RCEC cells were washed with PBS and incubated with annexin-V labeling solution (containing 10 μL annexin-V and 5 μL PI solution in 200 μL incubation buffer) for 15 min in the dark. Confocal images were acquired on a Nikon A1R Eclipse Ti confocal microscope (Nikon Corporation, Tokyo, Japan).
7
Quantitative Apoptosis Assay in FHC Cells
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Briefly, 1 × 105 treated FHC cells were attained. Apoptosis was subsequently detected using an Annexin V-fluorescein isothiocyanate/propidium iodide (PI) apoptosis detection kit (Beyotime, Shanghai, China), and the stained cells were identified with a flow cytometer. Annexin V-positive and PI-negative cells were regarded as apoptotic cells. The apoptosis rate was calculated using FlowJo software (Treestar, San Carlos, CA).
8
Annexin V-FITC/PI Apoptosis Assay
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Apoptosis was detected with the Annexin V‐fluorescein isothiocyanate/propidium iodide (PI) Apoptosis Detection Kit (Beyotime) according to the manufacturer's instructions. Briefly, cells were digested with trypsin and collected by centrifugation at 300g for 5 min. After washing twice with phosphate buffer saline, 100 µl binding buffer was added to resuspend the cells. Then, the cells were incubrated with 5 μl Annexin V and 10 µl PI for 15 min at room temperature in the dark. Finally, the cells were detected using a flow cytometer (BD Biosciences).
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