Cell apoptosis assay was performed by flow cytometric analysis using an annexin V-fluorescein isothiocyanate kit (BD Biosciences, Germany). In brief, cells were kept in serum-free medium for 16 h, followed by harvesting with ice-cold phosphate-buffered solution and re-suspending with binding buffer. Then, cells (5×106 cells/well) were stained with annexin V-fluorescein isothiocyanate (0.5 μg/mL) and propidium iodide (0.6 μg/mL) for 15 min in the dark at room temperature, and immediately analyzed on a FACS CaliburTM system (Becton Dickinson, USA).
Annexin 5 fluorescein isothiocyanate kit
Manufactured by BD
Sourced in Germany
The Annexin V-fluorescein isothiocyanate (FITC) kit is a laboratory reagent used for the detection and quantification of apoptosis (programmed cell death) in cell samples. The kit contains Annexin V, a protein that binds to phosphatidylserine, a molecule exposed on the surface of apoptotic cells. Annexin V is conjugated with the fluorescent dye FITC, allowing for the visualization and analysis of apoptotic cells using flow cytometry or fluorescence microscopy.
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2 protocols using annexin 5 fluorescein isothiocyanate kit
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Annexin V-FITC Cell Apoptosis Assay
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Annexin V-FITC Apoptosis Assay
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The Annexin V Fluorescein Isothiocyanate kit (BD Pharmingen, San Diego, CA) was used to detect cell apoptosis. The cells were collected after 48hrs transfection with miRNA mimic or gene construct. After then, cells were suspended in 100 ml binding buffer (1 × 106 cells/ml) and incubated with Annexin V-FITC and propidium iodide for 15 min. The apoptosis was detected on a BD FACSCalibur™ system (Becton-Dickinson, Franklin Lakes, NJ, USA). miR-NC or VC were used as the negative controls for miRNA or gene, respectively.
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