Approximately 1 × 105 cells of MCF-10A-Tr were grown and treated as above before harvest and experiment was carried out according to the Annexin V-FITC/PI detection kit protocol (Sigma). Cells were sorted using Flow cytometer (Attune NxT Flow Cytometer) at an event count of 10,000 cells/sample. The data obtained was analyzed by using FCS express 5 software.
Annexin 5 fitc pi detection kit
Manufactured by Merck Group
Sourced in United States
The Annexin V-FITC/PI detection kit is a laboratory product designed to detect and quantify apoptosis, a process of programmed cell death, in cell samples. The kit utilizes Annexin V, a protein that binds to phosphatidylserine, and propidium iodide (PI), a DNA-binding dye, to differentiate between viable, early apoptotic, and late apoptotic/necrotic cells. The kit provides a straightforward method for analyzing cellular viability and the stage of apoptosis through flow cytometry or fluorescence microscopy.
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5 protocols using annexin 5 fitc pi detection kit
1
Apoptosis Assay in MCF-10A-Tr Cells
2
Apoptosis Analysis of HK-2 Cells
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HK-2 cells after various treatments were washed five times with ice-cold PBS. The cells (1.0 × 105) were resuspended in 200 µL of binding buffer containing 2 µL of Annexin V-fluorescein isothiocyanate (FITC) and 4 µL of propidium iodide (PI), referring to the suggestion of the Annexin V-FITC/PI Detection Kit (Sigma-Aldrich). The apoptotic cells were analyzed by a flow cytometer (BD Bioscience, Stockholm, Sweden) with a 488 nm laser.
3
Annexin V-FITC/PI Apoptosis Assay
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The apoptosis of HPMVECs were determined by flow cytometry using an Annexin V-FITC/PI detection kit (Sigma-Aldrich; Merck KGaA) in accordance with a previous study (30 (link)). Briefly, cells were washed with PBS, re-suspended in binding buffer and stained with Annexin V/FITC and PI solution for 30 min. The apoptotic cells were detected with a flow cytometer (Beckman Coulter, Inc.) and Expo32 v1.2 software (Beckman Coulter, Inc.).
4
Quantifying Apoptosis by Flow Cytometry
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Cell apoptosis was investigated by flow cytometry using the Annexin V-FITC/PI detection kit (Sigma-Aldrich; Merck KGaA). Following treatment with PTX for 24 h, Huh7 and Huh7/PTX were resuspended in binding buffer and then incubated with 10 µl Annexin V-FITC and PI for 20 min at room temperature in the dark. The positive cells (Annexin V-FITC+/PI−/+) were examined using a flow cytometer (BD Biosciences) and BD FACStation software (version 6.1; BD Biosciences)
5
Flow Cytometric Analysis of Apoptosis
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Cardiomyocytes were seeded onto 12-well plates at a density of 1 × 105 cells/well. After treatment as specified in the Results section, the cells were trypsinized and harvested for staining using the Annexin V-FITC/PI Detection Kit, according to the manufacturer’s instructions (Sigma-Aldrich, St. Louis, MO, USA). Cells were analysed by flow cytometry (Becton-Dickinson, Franklin Lakes, NJ, US). The FITC + /PI− fraction and FITC + /PI + fraction were considered apoptotic cells (early and late apoptosis, respectively).
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