For the cell DNA replication assay, we pre-cultured primary phGC and poGC for 16 h in a 96-well plate at a density of 50,000 cells/well. Then, we transfected the phGC and poGC with FOXL2-siRNA or NC-siRNA. Cell DNA replication was studied in each well utilizing a Cell Light EdU DNA imaging kit (RiboBio, China) following the manufacturer’s protocol. Briefly, fresh culture medium with 5-ethynyl-2’-deoxyuridine (10 μM EdU from the Cell Light EdU DNA imaging kit, Guangzhou RiboBio, China) was used as the replacement medium 6 h post-transfection and then cultured for 24 h. Finally, the cell nuclei were re-stained with Hoechst 33342 and observed under a fluorescence microscope (Eclipse, Nikon, Japan). EdU-positive cells were regarded as cell DNA replication-positive, the number of which were calculated as (EdU add-in cells/Hoechst stained cells) ×100%.
Cell light edu dna imaging kit
Manufactured by RiboBio
Sourced in China
The Cell Light EdU DNA imaging Kit is a lab equipment product designed for DNA imaging and detection. It provides a method for visualizing and analyzing DNA incorporation during cell proliferation. The kit contains essential components for labeling, detecting, and analyzing DNA in cells.
Automatically generated - may contain errors
Lab products found in correlation
16 apollo reaction cocktail, by RiboBio (2 mentions)
High content imaging pathway 855, by BD (2 mentions)
Inverted microscopy, by Nikon (2 mentions)
Hoechst 33342, by Merck Group (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Fluorescence microscopy, by Zeiss (1 mentions)
Molecular imager chemidoc xrs system, by Bio-Rad (1 mentions)
Inverted florescence microscope, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by RiboBio (1 mentions)
Tcs sp8 confocal microscope system, by Leica (1 mentions)
Synergy microplate reader, by Agilent Technologies (1 mentions)
Cck 8 solution, by Beyotime (1 mentions)
Spectramax 250, by Molecular Devices (1 mentions)
Fsx100 microscope, by Olympus (1 mentions)
Modfit lt 5, by Verity Software House (1 mentions)
Cell cycle detection kit, by Keygen Biotech (1 mentions)
Spectramax i3x microplate reader, by Molecular Devices (1 mentions)
Cck 8 reagent, by Dojindo Laboratories (1 mentions)
Cck 8 assay kit, by Dojindo Laboratories (1 mentions)
38 protocols using cell light edu dna imaging kit
1
Quantifying DNA Replication in Primary Granulosa Cells
2
Quantifying Cell DNA Synthesis
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To determine the DNA synthesis, the Cell Light EdU DNA imaging kit (RiboBio Co., Guangzhou, China) was used. In short, cells were seeded in 24-well plates and exposed to EdU for 2 h. Then they were fixed in 4% paraformaldehyde and permeabilized in 0.5% Triton X-100. Images were taken using a fluorescent microscope at 488 nm excitation. Each experiment was repeated independently thrice.
3
EdU Assay for Measuring Cell Proliferation
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An EdU assay was performed using the Cell Light EdU DNA imaging kit (Guangzhou RiboBio Co., Ltd.)to measure the effects of let-7g/i on cellular proliferation. The EdU assay was performed 48 h after cells were transfected with let-7g/i agomir. Cells were seeded in 96-well plates and exposed to 25 mm EdU for 2 h at 37°C, and were then fixed in 4% paraformaldehyde. Following permeabilization with 0.5% Triton X-100 (Amresco LLC, Solon, OH, USA), the 16 Apollo reaction cocktail (Guangzhou RiboBio Co., Ltd.) was added and the cells were incubated for 30 min. Subsequently, the DNA of the cells was stained with Hoechst 33342 (Sigma-Aldrich, St. Louis, MO, USA) for 30 min and visualized under a fluorescent microscope (IX81; Olympus Corporation, Tokyo, Japan). The cell count was analyzed by Image-Pro Plus 6.0 software (Media Cybernetics, Inc., Rockville, MD, USA).
4
EdU Proliferation Assay for Lung Cancer
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The EdU proliferation assay was used to test lung cancer cell proliferation according to the Kit protocol (Cell Light EdU DNA imaging Kit, Guangzhou RiboBio, China). In brief, A549 and 95D cells were transfected with miRNA mimics in 96-well plates. Forty-eight hours after transfection, 50 mM 5-ethynyl-2′-deoxyuridine (EdU) was added and the cells were cultured for an additional 2 hours. The cells were then stained according to the Kit protocol: discard the EdU medium mixture, wash cells with PBS, add 4% paraformaldehyde (PFA) to fix cells at room temperature for 30 minutes, wash with glycine (2 mg/ml) for 5 minutes in a shaker, add 0.2% Trion X-100 for 10 minutes, wash with PBS for two times, add click reaction buffer (Tris–HCl, pH 8.5, 100mM; CuSO4, 1mM; Apollo 550 fluorescent azide,100 mM; ascorbic acid, 100mM) for 30 minutes while protecting from light, wash with 0.5% Triton X-100 for three times, stain with DAPI (0.5 mg/L) for 30 minutes at room temperature, wash with 0.5% Triton X-100 for five times, and finally add 150 ul PBS. Images were taken and analyzed using High Content Imaging Pathway 855 (BD, USA). EdU positive cells were calculated with EdU% = (EdU-add-in cells/Hoechst stained cells)∗ 100%.
5
Evaluating Proliferation and Invasion in Prostate Cancer
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First, a cell counting kit-8 (CCK-8; Dojindo, Kumamoto, Japan) and a 5-ethynyl-20-deoxyuridine assay (EdU) kit (Cell Light EdU DNA imaging Kit, RiboBio, Guangzhou, China) were used to evaluate the proliferative activity of PC cells according to the manufacturer’s instructions. Second, colony formation assays were conducted to evaluate the long-term proliferation of PC cells as described in our previous study [29 (link)]. Third, wound-healing assays were carried out by generating a vertical scratch on a monolayer of PC cells. Images were captured under an inverted microscope after 24 h, and the wound area was calculated in five randomly selected microscopic fields. Additionally, transwell chamber assays were performed to assess cell invasive ability in accordance with the method described in our previous study [29 (link)].
6
Cell Proliferation Assay with EdU
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An EdU assay was carried out using a Cell Light EdU DNA imaging kit (RiboBio). The ratio of EdU‐stained cells (with red fluorescence) to Hoechst‐stained cells (with blue fluorescence) was used to examine cell proliferation.
7
Quantifying Glioma Cell Proliferation
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We performed the EdU assay using Cell Light EdU DNA imaging Kit (RiboBio Co., Ltd. Guangzhou, China) to analyze cell proliferation. C6 glioma cells were incubated with EdU for 24h after appropriate treatments with different concentrations of GDNF. The percentage of EdU+ C6 cells was determined from the images.
8
Evaluating Cell Proliferation with EdU
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An EdU kit (Cell Light EdU DNA imaging kit, RiboBio, Guangzhou, China) was used to evaluate cell proliferation according to the manufacturer's instructions. Images were detected and analyzed with a microscope (Olympus, Tokyo, Japan). The average ratio of EdU-stained cells (red) to DAPI-stained cells (blue) was used to evaluate cell proliferation activity.
9
Quantifying Cell Proliferation via EdU Assay
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The cells were incubated with 5-ethynyl-2-deoxyuridine (EdU) (100 μM, Cell Light EdU DNA imaging Kit, Guangzhou RiboBio, China) for an additional 6 h. Subsequently, cell staining was conducted according to standard protocol. Briefly, after discarding the EdU medium mixture, 4% paraformaldehyde was added to fix cells for 30 min at room temperature, followed by washing with glycine (2 mg/mL) for 5 min. After addition of 0.2% Trion X-100 for 10 min, the cells were washed with PBS twice, followed by addition of click reaction buffer for 10–30 min in the dark. Then, the cells were washed with 0.5% Triton X-100 for three times, and stained with Hoechst33342 (5 μg/ml) at room temperature for 30 min. After washing with 0.5% Triton X-100 for five times, 150 μL PBS was added to the wells. EdU staining images were photographed under fluorescent microscope (Nikon, Ti-s), and the proportion of EdU positive cells was obtained using the formula: (EdU add-in cells/Hoechst stained cells) × 100%.
10
Lycorine's Impact on Cell Proliferation
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MNNG/HOS and U2OS cells were seeded in 96-well plates at a concentration of 3×103 cells per hole and treated with various concentrations of Lycorine for48 h. Then 100 μl of 50 μM EdU (Cell Light EdUDNA Imaging Kit; Guangzhou RiboBio, China) was added to each well, and the cells were cultured in the medium for an additional 2 h. The cells were then stained as previously described.25 (link) Finally, the cells were observed under fluorescence microscopy and images were taken. EdU positive cells were calculated with the following formula: (EdU add-in cells/Hoechst stained cells) × 100%.26 (link)
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