Transfected CM cells were treated with EdU medium diluent (Ribobio) for 3 hours, then with 4% paraformaldehyde and 0.5% Troxin X‐100. After staining with Apollo® 488 fluorescent at 37°C for 30 minutes, DAPI (Beyotime) was utilized for nuclear staining, followed by visualization under fluorescent microscope (Leica). Three repeated EdU assays were conducted indistinguishably to assess the proliferation of different cells.
Edu medium diluent
Manufactured by RiboBio
Sourced in China
EdU medium diluent is a buffered solution used to dilute EdU (5-Ethynyl-2'-deoxyuridine) reagent prior to cell labeling experiments. It is designed to maintain the stability and activity of the EdU compound in cell culture media.
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Lab products found in correlation
3 protocols using edu medium diluent
1
Cell Proliferation Quantification via EdU
2
Quantification of Cellular Proliferation
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LX2 cells were incubated with 50 mM EDU medium diluent (RiboBio, Guangzhou, China) in 96-well plates for 3 h. Following being fixed and permeabilized, 1 × Apollo® reaction cocktail (100 μL) was employed to react with the EdU for 30 min. Cell nuclei were stained with DAPI. Finally, EdU positive cells were imaged by the fluorescence microscopy (Bio-Rad, Hercules, CA) and analyzed by ImageJ software (Bethesda, MD).
3
Cell Proliferation Assays for OS Cells
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Cell counting kit-8 (CCK-8) assays were used to analyze OS cell proliferation. 143B and U2OS cells were put into 96-well plates and cultured with 10 μl/well CCK-8 (Beyotime, Jiangsu, China) solution for 48 h. The cellular absorbance was detected by microplate reader (Molecular Devices) at 450 nm. For EdU incorporation assay, cells were transfected and incubated with 100 μL of 50 μM EdU medium diluent (Ribobio, Guangzhou China) in culture plates for 3 h. After xation, cells were treated with 100 μL of the 0.5% Troxin X-100 and 100 μL of 1 × Apollo ® 488 uorescent staining reaction liquid for 30 min at 37℃. Cell nuclei were dyed with DAPI. All proliferative assays were repeated for at least three times.
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