Cell apoptosis analysis was carried out with the help of PI and FITC‐conjugated Annexin V staining (Beyotime). In short, after transfection, cells were collected and centrifuged and then were resuspended in 195 μL of Annexin V‐FITC binding buffer. Afterward, 5 μL of Annexin V‐FITC and 10 μL of PI were, respectively, added and reacted at 25°C in dark condition for 15 minutes. Thereafter, the results were measured by utilizing FACS can (Beckman Coulter). The data were analyzed by FlowJo software (Tree Star Software).
Fitc conjugated annexin 5 staining
Manufactured by Beyotime
Sourced in China
FITC-conjugated Annexin V is a fluorescent-labeled protein used to detect and quantify apoptosis (programmed cell death) in cell populations. Annexin V is a calcium-dependent phospholipid-binding protein that has a high affinity for phosphatidylserine, which is exposed on the outer membrane of apoptotic cells. The FITC (Fluorescein Isothiocyanate) label allows for the detection of Annexin V binding through fluorescence microscopy or flow cytometry.
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2 protocols using fitc conjugated annexin 5 staining
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Apoptosis Evaluation by Annexin V-FITC/PI Staining
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Annexin V and PI Apoptosis Assay
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Cell apoptosis analysis was performed using propidium iodide (PI) and fluorescein isothiocynate (FITC)-conjugated Annexin V staining (Beyotime Biotechnology, Shanghai, China). Briefly, cells were washed in phosphatebuffered saline (PBS) and fixed in 70% ethanol. Fixed cells were then washed twice in PBS and stained in PI/FITC-Annexin V in the presence of 50 μg/ml RNase A (Sigma-Aldrich), and then incubated for 1 h at room temperature in the dark. Flow cytometry analysis was done by using a FACS can (Beckman Coulter, Fullerton, CA, USA). The data were analyzed by using FlowJo software.
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