Flow cytometric assessment of cell viability was performed using the FITC-annexin V/propidium iodide assay (Thermo Fisher Scientific). HaCaT cells were cultured in six-well plates and treated with ENDS for 24 hours, 48 hours, or 72 hours. Cells were detached by incubation with trypsin-EDTA and centrifuged at 1,500 rpm for 5 minutes. The cell pellet was collected and resuspended in 100 μL Annexin-Binding Buffer. FITC-annexin V and propidium iodide were then added to the cell suspension. After incubation at room temperature for 15 minutes, the stained cells were analyzed by flow cytometry (FACScan flow cytometer; BD, Franklin Lakes, NJ, USA), measuring the fluorescence emission at wavelengths of 530 nm and >575 nm.
Fitc annexin 5 propidium iodide assay
Manufactured by Thermo Fisher Scientific
Sourced in United States
The FITC-annexin V/propidium iodide assay is a laboratory tool used to detect and quantify apoptosis, a type of programmed cell death. It utilizes the binding properties of annexin V, a protein that has a high affinity for phosphatidylserine, which is externalized during apoptosis. The assay also incorporates propidium iodide, a DNA-binding dye that can penetrate cells with compromised membranes, indicative of late-stage apoptosis or necrosis. This combination of fluorescent markers allows for the differentiation of viable, early apoptotic, and late apoptotic/necrotic cells.
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Lab products found in correlation
2 protocols using fitc annexin 5 propidium iodide assay
1
Cell Viability Evaluation by Flow Cytometry
2
Quantitative Assessment of Cell Viability and Apoptosis
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Flow cytometric assessments of cell viability and apoptosis were determined with a FITC-annexin V/propidium iodide assay (Thermo Fisher Scientific, Waltham, MA, United States). Briefly, growing U87 and U118 cells were digested with 0.25% trypsin and counted. These cells were then diluted to a final concentration of 1 × 105 cells/ml and inoculated in a culture dish at a density of 10 mL/dish. The cells were then treated with artocarpin for the indicated times. Cells were detached by incubation with trypsin-EDTA and centrifuged at 1,500 rpm for 5 min. The pellet was collected and resuspended in 100 μL annexin-binding buffer. FITC-annexin V and propidium iodide were then added to the cell suspension. After incubation at room temperature for 15 mins, the stained cells were analyzed by flow cytometry (FACScan; Becton Dickinson, Franklin Lakes, NJ, United States). Fluorescence emission was measured at λ = 530 nm (excitation) and at λ = 575 nm (emission).
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