The cells were routinely infected, cultured for 48 hr, and then digested with trypsin. An Annexin V‐FITC/propidium iodide (PI) Apoptosis Assay Kit (Biyuntian Biotechnology Co. Ltd.) was used to estimate the apoptotic rate according to the manufacturer's instructions. The cells were suspended in 1 × Annexin V‐binding buffer, and then 5 μl of Annexin V and 1 μl of PI were added to 100 μl of cell suspension and mixed. The mixture was incubated in the dark for 15 min at room temperature, after which 400 μl of the 1 × Annexin V‐binding buffer was added to each sample to terminate the reaction. The rate of apoptosis was determined using a FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ).
Annexin 5 fitc propidium iodide apoptosis assay kit
Manufactured by Beyotime
The Annexin V-FITC/propidium iodide (PI) Apoptosis Assay Kit is a laboratory reagent used to detect and quantify apoptosis, a form of programmed cell death. The kit utilizes Annexin V, a calcium-dependent phospholipid-binding protein, conjugated with the fluorescent dye FITC, and the DNA-binding dye propidium iodide. This combination allows for the identification of early apoptotic cells, late apoptotic/necrotic cells, and viable cells through flow cytometry or fluorescence microscopy analysis.
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Annexin V-FITC/PI Apoptosis Assay
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Annexin V-FITC/PI Apoptosis Assay
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Trypsin was used to digest cells after culturing for 48 h. The apoptotic rate was assessed using Annexin V FITC/propidium iodide (PI) Apoptosis Assay Kit (Biyuntian Biotechnology Co. Ltd.) as per the manufacturer’s instructions. The cells were suspended in 1 × Annexin V binding buffer and Annexin V (5 μL) and PI (1 μL) were added to the cell suspension (100 μL) and mixed. The mixture was incubated in darkness at room temperature for 15 min, and then 1 × Annexin V binding buffer (400 μL) was added the each sample to stop the reaction. The apoptosis rate was measured using FACSCalibur flow cytometer (BD Biosciences, Franklin Lakes, NJ).
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