To quantify apoptotic cells, flow cytometry was performed with an Annexin V-fluorescein-5-isothiocyanate apoptosis detection kit (Bio-vision, USA). After transfection for 48 h, cells were harvested in a 5-mL tube. Then, the cells were washed with cold PBS and re-suspended in 1×binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4) at a final concentration of 1×106 cells/mL. FITC-AnnexinV (5 μL) and propidium iodide (PI, 5 μL) were gently mixed and incubated with the cells for 15 min at room temperature. After incubation, the samples were analyzed by flow cytometry within 1 h.
Annexin 5 fluorescein 5 isothiocyanate apoptosis detection kit
Manufactured by Abcam
Sourced in United States
The Annexin V-fluorescein-5-isothiocyanate apoptosis detection kit is a laboratory tool used to detect and quantify the level of apoptosis, or programmed cell death, in samples. It utilizes Annexin V, a protein that binds to phosphatidylserine, a molecule that is exposed on the surface of cells undergoing apoptosis. The kit includes Annexin V conjugated to the fluorescent dye fluorescein-5-isothiocyanate (FITC), which allows for the visualization and measurement of apoptotic cells using techniques such as flow cytometry or fluorescence microscopy.
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2 protocols using annexin 5 fluorescein 5 isothiocyanate apoptosis detection kit
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Quantifying Apoptosis via Flow Cytometry
2
Annexin V-based Apoptosis Quantification
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To quantify the apoptotic cells, flow cytometry was performed using an Annexin V-fluorescein-5-isothiocyanate Apoptosis Detection kit (BioVision, Inc., Milpitas, CA, USA). Forty-eight hours after transfection with miR-518a-3p mimics (50 nM) or negative control (50 nM), the LoVo cells were placed in a 5-ml tube. The cells were then washed with cold PBS and resuspended at a final concentration of 1×106 cells/ml. FITC-Annexin V (5 μl) and propidium iodide were gently mixed and incubated with the cells for 15 min at room temperature. Within 1 h after incubation, the samples were analyzed by flow cytometry.
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