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5 ethynyl 2 deoxyuridine (edu)

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EdU is a nucleoside analog of thymidine that is incorporated into newly synthesized DNA during the S phase of the cell cycle. It can be used to label and detect proliferating cells in various biological applications.

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Lab products found in correlation

5 ethynyl 2 deoxyuridine (edu), by RiboBio (6 mentions) Fluorescence microscope, by Nikon (6 mentions) Apollo reaction cocktail, by RiboBio (4 mentions) Fluorescence microscopy, by Nikon (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Cell light edu apollo 488 in vitro imaging kit, by Thermo Fisher Scientific (1 mentions) Az100 macro confocal microscope, by Nikon (1 mentions) Click it edu alexa fluor 488 imaging kit, by Thermo Fisher Scientific (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by Thermo Fisher Scientific (1 mentions) Hoechst 33342, by Beyotime (1 mentions) 80i microscope, by Nikon (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by Beyotime (1 mentions) Confocal microscope, by Nikon (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) Te2000 u, by Nikon (1 mentions) Te2000 microscope, by Nikon (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions)

13 protocols using 5 ethynyl 2 deoxyuridine (edu)

1

EdU Proliferation Assay Protocol

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For this assay, 10 μM 5-ethynyl-2ʹ-deoxyuridine (EdU, RiboBio, Guangzhou, Guangdong, China) was added into the growth medium and incubated for 3 h. Fixation, permeabilization, and EdU staining were done according to the manufacturer’s protocol. Cell nuclei were counter-stained with Hoechst 33,342 (Invitrogen, Carlsbad, CA, USA) at a concentration of 5 μg/ml for 10 min. Then, EdU-positive cells were visualized under a fluorescence microscope (Nikon, Tokyo, Japan) to calculate the ratio of EdU-positive cells (EdU-stained cells/total cells) [10 (link)].
Wu W., Xu K., Li M., Zhang J, & Wang Y. (2021). MicroRNA-29b/29c targeting CTRP6 influences porcine adipogenesis via the AKT/PKA/MAPK Signalling pathway. Adipocyte, 10(1), 264-274.
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2

Cell Proliferation Assay with EdU

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Cells were seeded in 48-well plates for 24 hours and incubated in medium containing 5-ethynyl-2-deoxyuridine (EdU) (50 mM; RiboBio Co., Guangzhou,China) for 2 hours. Cell fixation, staining, and DNA staining were performed according to the manufacturer’s instructions. Nuclei and EdU-positive cells were observed under a fluorescence microscope (Nikon, Tokyo, Japan), and the proliferation rate (EdU-positive cells/nucleus) was calculated in random fields.
Zhang W., Liu B., Wang Y., Sun PHD L., Liu C., Zhang H., Qin W., Liu J., Han L, & Shan W. (2022). miR-195-3p/BDNF axis regulates hypoxic injury by targeting P-ERK1/2 expression. Medicine, 101(46), e31586.
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3

Quantifying Cell Proliferation with EdU

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Transfected cells were incubated for 2 h with 50 μm EdU (RiboBio, Guangdong, China), followed by staining with DAPI (Invitrogen). The images were then captured by fluorescence microscopy (Nikon, Tokyo, Japan), and EdU‐positive cells were visualized and counted. The ratio of the total number of EdU‐positive cells to the total number of DAPI chromogenic cells was taken as the EdU‐positive rate.
Wu Q., Ma J., Wei J., Meng W., Wang Y, & Shi M. (2020). FOXD1‐AS1 regulates FOXD1 translation and promotes gastric cancer progression and chemoresistance by activating the PI3K/AKT/mTOR pathway. Molecular Oncology, 15(1), 299-316.
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4

EdU Incorporation Assay for Proliferation

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At 48 h after trans-fection, MPMCs were exposed to 10 mM EdU (Guangzhou RiboBio Co., Ltd., Guangzhou, China) for 24 h at 37°C. The cultured cells were then fixed with 4% paraformalde-hyde (PFA) for 30 min at room temperature and permeabilized with 0.5% Triton X-100. Next, the 1X Apollo reaction cocktail (Guangzhou RiboBio Co., Ltd.) was added to the cells and incubated for 30 min at room temperature. The cells were then stained using a Cell-Light™ EdU Apollo®488 in vitro imaging kit (Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. EdU-stained cells were observed using a fluorescence microscope (magnification, ×400; Nikon Corporation, Tokyo, Japan).
Han T., Wu N., Wang Y., Shen W, & Zou J. (2019). miR-16-2-3p inhibits cell proliferation and migration and induces apoptosis by targeting PDPK1 in maxillary primordium mesenchymal cells. International Journal of Molecular Medicine, 43(3), 1441-1451.
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5

EdU Labeling and Imaging Protocol

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For injection, EdU (Carbosynth Limited) was resuspended at 5 mg/mL in saline and passed through a 0.2 μm filter. One mg was given by intraperitoneal (IP) injection. For continuous administration, mice were given an IP injection at the start of the experiment and then provided with EdU in the water, resuspended at 0.5 mg/mL with 1% sucrose and passed through a 0.2 μm filter. Water bottles were shielded from light and the water supply changed every three days. Samples were dissected and processed as for IF. For EdU detection, the Click-iT EdU Alexa Fluor 488 Imaging Kit (ThermoFisher Scientific) was used. For combined EdU and protein detection, EdU detection was completed first and then the IF protocol was followed beginning at the blocking step. Whole-mounts in which EdU was detected were acquired in a single frame with z-stack on the Nikon AZ100 macro confocal microscope and processed, including with maximum projection, with Fiji (ImageJ). Sections were imaged as described above.
Frumm S.M., Yu K.S., Chang J., Artichoker J.A., Scaria S.M., Lee K.P., Byrnes L.E., Sneddon J.B, & Tward A.D. (2020). A hierarchy of proliferative and migratory keratinocytes maintains the tympanic membrane. Cell stem cell, 28(2), 315-330.e5.
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6

HCC Cell Proliferation Assay

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EdU incorporation assays were used to measure HCC cell proliferation. Briefly, HCC cells (1 × 105 cells/well) were grown in dishes and treated for 48 h. Cells were then labeled with 50 μM EdU (Beyotime Institute of Biotechnology, Shanghai, China) for 2 h at 37°C. Cells were subsequently fixed in 4% PFA for 30 min, permeabilized with 0.5% Triton X-100 for 30 min and incubated with Apollo reaction cocktail (500 μL/well) for 30 min. Cell nuclei were Hoechst 33342 (Beyotime Institute of Biotechnology, Shanghai, China) stained and cells were imaged via fluorescence microscopy (10 fields of view per treatment). EdU-positive cells were counted on a Nikon, 80i microscope (Nikon, Japan). Average EdU intensities per cell were measured.
Liu S., Qiu J., He G., Geng C., He W., Liu C., Cai D., Pan H, & Tian Q. (2020). Dermatopontin inhibits WNT signaling pathway via CXXC finger protein 4 in hepatocellular carcinoma. Journal of Cancer, 11(21), 6288-6298.
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7

Adipocyte Proliferation Assay with EdU

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This cellular proliferation assay involved fluorescence detection using EdU (RuiBo, Guangzhou, China), a thymidine analog that can be incorporated into newly synthesized DNA during cell proliferation. At 24 h post-transfection, adipocytes were incubated with EdU (50 μM) for 2 h, fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and labeled with Apollo® fluorescent dye (RuiBo, Guangzhou, China) following the manufacturer’s protocol. The EdU-positive cells were imaged with a confocal microscope (Nikon, Tokyo, Japan) to calculate their percentage.
Zhang J., Yang J., Yang N., Ma J., Lu D., Dong Y., Liang H., Liu D, & Cang M. (2020). Dlgap1 negatively regulates browning of white fat cells through effects on cell proliferation and apoptosis. Lipids in Health and Disease, 19, 39.
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8

Quantifying Adipocyte Proliferation

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After transfection for 48 h, intramuscular adipocytes were incubated at 37°C with 50 μM EdU (RiboBio, China) for 2 h, then cells were fixed with 4% PFA for 30 min and neutralized by 2 mg/mL glycine solution, permeabilized with 0.5% Triton X-100. Then cells were incubated with Apollo Reaction Cocktail (RiboBio, China) for 30 min at room temperature. The DNA was stained with DAPI (Beyotime) for 15 min. The EdU-positive cells were observation with a fluorescence microscope (Nikon, Tokyo, Japan).
Zhang M., Li D., Zhai Y., Wang Z., Ma X., Zhang D., Li G., Han R., Jiang R., Li Z., Kang X, & Sun G. (2020). The Landscape of DNA Methylation Associated With the Transcriptomic Network of Intramuscular Adipocytes Generates Insight Into Intramuscular Fat Deposition in Chicken. Frontiers in Cell and Developmental Biology, 8, 206.
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9

Cell Proliferation Analysis by EdU Assay

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At 48 h after transfection, DF-1 cells were incubated at 37°C for 2 h in the presence of 50 μM EdU (RiboBio, China). The cells were then fixed in 4% paraformaldehyde for 30 min and neutralized using 2 mg/mL glycine solution. The cells were permeabilized by adding 0.5% Triton X-100. A solution containing EdU (Apollo Reaction Cocktail; RiboBio, China) was added and the cells were incubated at room temperature for 30 min. The nuclear stain Hoechst 33342 was then added and incubation was continued for another 30 min. The number of EdU-stained cells was determined using images of three randomly selected fields obtained with a fluorescence microscope (TE2000-U; Nikon, Japan).
Cai B., Li Z., Ma M., Wang Z., Han P., Abdalla B.A., Nie Q, & Zhang X. (2017). LncRNA-Six1 Encodes a Micropeptide to Activate Six1 in Cis and Is Involved in Cell Proliferation and Muscle Growth. Frontiers in Physiology, 8, 230.
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10

Cell Proliferation Assay via EdU Staining

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The human SSC line was seeded in 96-well plate containing DMEM/F12 medium with 50 μM EDU (RiboBio, Guangzhou, China). After 12 hr of culture, the cells were washed with DMEM and fixed with 4% PFA. Cells were neutralized with 2 mg/mL glycine and permeabilized with 0.5% Triton X-100 for 10 min at room temperature. EDU immunostaining was performed with Apollo staining reaction buffer. The nuclei of cells were stained with Hoechst 33342, and the EDU-positive cells were counted under fluorescence microscopy (Nikon).
Fu H., Zhang W., Yuan Q., Niu M., Zhou F., Qiu Q., Mao G., Wang H., Wen L., Sun M., Li Z, & He Z. (2018). PAK1 Promotes the Proliferation and Inhibits Apoptosis of Human Spermatogonial Stem Cells via PDK1/KDR/ZNF367 and ERK1/2 and AKT Pathways. Molecular Therapy. Nucleic Acids, 12, 769-786.
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