EdU staining was performed using EdU Imaging Kits (Cy3) (APExBIO) according to the manufacturer’s protocol. Cells seeded on glass slides were prepared and exposed to a range of EdU concentrations (5, 10, and 20 μM) for 2 h at 37°C. Further, the cells were fixed with 4% paraformaldehyde for 15 min and permeabilized with 0.5% Triton X-100/PBS for 20 min at room temperature. For the EdU click reaction, the cells were treated with staining solution for 30 min at room temperature in the dark. Nuclei were stained with 0.2 mg/mL DAPI.
Edu imaging kits cy3
Manufactured by Apexbio
Sourced in United States
The EdU Imaging Kits (Cy3) from Apexbio are a set of reagents designed for the detection and visualization of proliferating cells. The kits utilize the incorporation of the thymidine analog EdU (5-ethynyl-2'-deoxyuridine) into the DNA of dividing cells, which can then be labeled with a Cy3 fluorescent dye for imaging and analysis.
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7 protocols using edu imaging kits cy3
1
Quantifying Cell Proliferation via EdU Staining
2
EdU Incorporation and Visualization Protocol
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EdU incorporation was performed using EdU Imaging Kits-Cy3 (APExBIO) according to the manufacturer’s protocols. Briefly, cells were incubated with 50 μM EdU for 2 h and then fixed in 4% paraformaldehyde for 15 min. Subsequently, cells were permeabilized with 0.5% Triton X-100 for 20 min, and stained with a click reaction cocktail for 30 min. Cell nuclei were stained with 5 μg/mL Hoechst 33342 for 5 min. The number of EdU-positive cells was counted and photographed under a microscope.
3
Quantifying Cellular Proliferation with EdU
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EdU Imaging Kits (Cy3) (APExBIO, Houston, Texas, USA) were used to evaluate the cellular proliferation. Briefly, Huh7 cells were seeded into 24-well plates at a density of 7×104 cells/well and incubated for 24 h. After re-incubation with the EdU reagent for 2h, the cells were fixed and stained for 30 min. Nucleic acid was stained with Hoechst 33342. Representative images were obtained using a fluorescence microscope.
4
Cell Proliferation Assay with EdU Imaging
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Cells were seeded in 6-well plates at a density of 1 × 106, with a total of 8 groups with 3 replicates. The experiment was performed according to the instructions of EdU Imaging Kits (Cy3) (Cat# K075, APExBIO), the prepared EDU reagent was added to the cell culture medium and incubated for 9 h at 37°C and 5% CO2, after that, the medium was removed, and fixation solution was added for 10 min in the dark, followed by permeabilization solution for 10 min, after staining the nuclei. A fluorescence microscope was used for observation.
5
EdU Imaging for Cell Proliferation
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According to the manufacturer's instructions for the EdU Imaging Kits (Cy3) (APExBIO, TX, USA), BEAS-2B cells at a density of 6 × 103 cells per well were incubated with EdU solution in 96-well plates after 24 h of CSE–LPS coexposure along with medicinal intervention [11 (link)]. Following cell fixation, membrane permeabilization, and click reaction in turn, DNA synthesis in the cells was examined using the ImageXpress Micro 4 Widefield High-Content Analysis System (Molecular Devices, Sunnyvale, CA, USA) [12 (link)] to assess cell proliferation activity. The experiment was repeated three times.
6
Assessing Cell Proliferation via EdU Imaging
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EdU Imaging Kits (Cy3) (APExBIO, Houston, Texas, USA) were used for assessing cell proliferation. HNSCC cells were seeded in a 24-well plate (2 × 104 cells per well) and incubated overnight. After re-incubation with the EdU reagent for 2 h, and after fixing and staining, nucleic acid was stained with Hoechst 33342. Images were acquired using a fluorescence microscope (Leica DMI6000B).
7
EdU Proliferation Assay in FaDu Cells
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For EdU analysis, FaDu cells were cultured with 10 µm EdU for 2 h and then treated with 4% paraformaldehyde for fixation. After permeabilization, EdU Imaging Kits (Cy3) (K1075, Apexbio) was used according to the manufacturer's instructions. Hoechst 33 342 solution was used to counterstain cell nuclei. Proliferative cells were determined under the confocal microscope.
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