To measure cell cycle distributions, cells were collected, washed with PBS, and fixed in 70% (v/v) ethanol overnight. Cells were digested with 100 mg/ml of RNase A in PBS and stained in the dark for 30 min with 40 mg/ml of propidium iodide (Sigma Aldrich), followed by flow cytometry analysis. To measure apoptosis, cells were collected, washed with PBS, and stained with fluorescein isothiocyanate-labeled annexin V (Invitrogen) and propidium iodide, followed by flow cytometry analysis.
Fluorescein isothiocyanate labeled annexin 5
Manufactured by Thermo Fisher Scientific
Sourced in United States
Fluorescein isothiocyanate-labeled annexin V is a fluorescently labeled protein used to detect and quantify apoptosis in cells. Annexin V binds to phosphatidylserine, which is exposed on the surface of cells undergoing apoptosis. The fluorescein isothiocyanate (FITC) label allows for the visualization and detection of annexin V-positive cells using fluorescence-based techniques.
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2 protocols using fluorescein isothiocyanate labeled annexin 5
1
Cell Cycle and Apoptosis Analysis
2
Annexin V Apoptosis Assay
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Cell death was assessed by annexin V/propidium iodide (PI) staining. Phosphatidylserine expression, as an early sign of apoptosis, was determined by fluorescence microscopy analysis through the binding of fluorescein isothiocyanate-labeled annexin V (Invitrogen, USA) to phosphatidylserine. PI was used to differentiate necrotic cells. Hoechst dye was used for nuclear staining. Fluorescence images were recorded using the IX-71-22FL/PH microscope (Olympus, Japan).
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