Select cultures were transduced to stably express H2B-GFP as an independent measure for proliferation. The GFP positive cells were selected using using BD FACSAria II cell sorter. The proliferation of the H2B-GFP cell lines was determined using IncuCyte live-cell imaging systems. The cell counts were determined based on the number of GFP-labelled nuclei and the data were exported to GraphPad Prism for statistical analysis and graph generation. The cell-cycle progression was determined using a chemiluminescent ELISA BrdU incorporation assay (Sigma; 11669915001) following the manufacturer’s protocol. Luminescence was read on a BioTEK Synergy 2 plate reader. Each drug treatment was performed in triplicates and confirmed in two independent experiments.
Elisa brdu incorporation assay
Manufactured by Merck Group
The ELISA BrdU incorporation assay is a laboratory technique used to measure cell proliferation. It detects and quantifies the incorporation of the thymidine analog bromodeoxyuridine (BrdU) into the DNA of dividing cells. The assay involves the detection of incorporated BrdU using an anti-BrdU antibody, providing a measure of DNA synthesis and cell proliferation.
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2 protocols using elisa brdu incorporation assay
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Proliferation Analysis of H2B-GFP Cells
2
Proliferation Analysis of H2B-GFP Cells
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Select cultures were transduced to stably express H2B-GFP as an independent measure for proliferation. The GFP positive cells were selected using using BD FACSAria II cell sorter. The proliferation of the H2B-GFP cell lines was determined using IncuCyte live-cell imaging systems. The cell counts were determined based on the number of GFP-labelled nuclei and the data were exported to GraphPad Prism for statistical analysis and graph generation. The cell-cycle progression was determined using a chemiluminescent ELISA BrdU incorporation assay (Sigma; 11669915001) following the manufacturer’s protocol. Luminescence was read on a BioTEK Synergy 2 plate reader. Each drug treatment was performed in triplicates and confirmed in two independent experiments.
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