Apoptosis was determined by the detection of nuclear morphology using ethidium bromide/acridine orange (EB/AO) staining [56 (link)]. KKU-100 and KKU-M156 cells (1 x 104 cells/well) were grown in 96-well plates at 37°C for 24 h and then treated with the studied compounds combined with doxorubicin at concentrations that showed a synergistic inhibitory effect on growth for 48 h. Cells treated with the same concentrations of the compounds or with doxorubicin alone were also examined. Cells cultured with 0.1% DMSO were used as controls. The treated cells were stained with an EB/AO mixture (Sigma Chemical, St. Louis, MO, USA) and observed under a Nikon fluorescent microscope. The percentage of apoptotic cells with condensed chromatin or fragmented chromatin was calculated from 500 counted cells.
Eb ao mixture
Manufactured by Merck Group
Sourced in United States
The EB/AO mixture is a laboratory reagent used for staining and visualizing cells or cellular components. It is a combination of two dyes: Ethidium Bromide (EB) and Acridine Orange (AO). This mixture is commonly used in various biological and biochemical applications that require the identification and analysis of nucleic acids, such as DNA and RNA.
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Lab products found in correlation
2 protocols using eb ao mixture
1
Apoptosis Quantification by EB/AO Staining
2
Apoptotic Nuclear Morphology Assessment
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Characteristic apoptotic nuclear morphological changes were determined by confocal fluorescent microscopy using ethidium bromide/acridine orange (EB/AO) staining. Ham-1 cells were treated with varying concentrations of forbesione or 5-FU, or their combination, for 24 h and then stained with 14 μl of 100 μg/ml of the EB/AO mixture (Sigma). The nuclear morphological of cells that were apoptotic (exhibiting condensed or fragmented chromatin) were assessed at 20x magnification.
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