The 5-ethynyl-2′-deoxyuridine (EdU) detection kit (Invitrogen) was used to evaluate the percentage of proliferating cells at the S phase. Briefly, 10 μmol/L EdU was added to the cells and cultured for 2 h at 37°C. The cells were fixed with 3.7% formaldehyde and washed with PBS. Then 0.5% Triton X-100 was added to the cells to increase the permeability of the cellular membrane. The cells protected from light were stained with 1 × Click-iT reaction cocktail for 30 min at room temperature. Hoechst 33342 dye was subsequently used to stain the cell nucleus following PBS washing. Images were captured under a confocal laser scanning microscope (Olympus, Center Valley, PA).
5 ethynyl 2 deoxyuridine edu detection kit
Manufactured by Thermo Fisher Scientific
The 5-ethynyl-2′-deoxyuridine (EdU) detection kit is a tool used to label and detect proliferating cells. It allows the visualization of cells undergoing DNA synthesis, a key indicator of cell division and proliferation.
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2 protocols using 5 ethynyl 2 deoxyuridine edu detection kit
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Quantifying Cell Proliferation with EdU
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Quantifying Cell Proliferation with EdU
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The 5-ethynyl-2′-deoxyuridine (EdU) detection kit (Invitrogen) was used to evaluate cell proliferation. According to the manufacturer's instructions, the cells were treated with 10 μmol/L EdU for 4 h at 37°C and fixed with 3.7% formaldehyde for 15 min. After cells were washed with 3% BSA in PBS, they were treated with 0.5% Triton X-100 (Sigma-Aldrich) for 20 min and stained with 1 × Click-iT reaction cocktail for 30 minutes at room temperature. After PBS wash, Hoechst 33342 dye was used to stain the cell nucleus at room temperature for 30 min. Images were captured under a confocal laser scanning microscope (Olympus, Center Valley, PA). The assay was repeated in triplicate.
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