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5 ethynyl 2 deoxyuridine (edu)

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EdU is a nucleoside analog that can be incorporated into DNA during DNA synthesis. It can be used to label and detect proliferating cells in various applications.

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Lab products found in correlation

5 ethynyl 2 deoxyuridine (edu), by Thermo Fisher Scientific (16 mentions) Bromodeoxyuridine (brdu), by Merck Group (12 mentions) Thymidine, by Merck Group (6 mentions) Hydroxyurea, by Merck Group (5 mentions) Click it edu imaging kit, by Thermo Fisher Scientific (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions) Fluorescence microscope, by Olympus (4 mentions) Mirin, by Merck Group (4 mentions) Auto 488, by Merck Group (4 mentions) Gallios flow cytometer, by Beckman Coulter (4 mentions) Doxycycline, by Merck Group (4 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (4 mentions) Click it plus edu flow cytometry assay kit, by Thermo Fisher Scientific (3 mentions) Hoechst 33342, by Merck Group (3 mentions) Cck 8, by Dojindo Laboratories (3 mentions) Tamoxifen, by Merck Group (3 mentions) Dimethyl sulfoxide (dmso), by Merck Group (3 mentions) 5 bromo 2 deoxyuridine, by Merck Group (2 mentions) Sodium ascorbate, by Merck Group (2 mentions)

Close spelling variants of this product

5-Ethynyl-2’- deoxyuridine (EdU, (2 citations) EdU (5-ethynyl-2’-deoxyuridine) proliferation assay (2 citations)

163 protocols using 5 ethynyl 2 deoxyuridine (edu)

1

Measuring eosinophil half-life in the gut

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Mice received 1 mg/ml EdU (Sigma-Aldrich) in drinking water for 3 or 6 d. EdU was replaced every 3 d. Eosinophil half-life (t1/2) in the small intestine and colon was calculated as t1/2 = t/log0.5(Nt/N0), where t is the time of EdU administration (6 d), Nt is the percentage of EdU– cells after 6 d, and N0 is the percentage of EdU– cells at time 0 (100%).
Diny N.L., Schonfeldova B., Shapiro M., Winder M.L., Varsani-Brown S, & Stockinger B. (2022). The aryl hydrocarbon receptor contributes to tissue adaptation of intestinal eosinophils in mice. The Journal of Experimental Medicine, 219(4), e20210970.
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2

Cardiac Tissue Sampling and Processing

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All mice were maintained and studied using protocols approved by the Institutional Animal Care and Use Committee (IACUC) of National University of Singapore. Animal work was undertaken in accordance with Singapore National Advisory Committee for Laboratory Animal Research guidelines. Relevant national and institutional guidelines and regulations must be consulted before commencement of any animal work. All studies were conducted in male C57BL/6JINV (Jax) mice. For virus injection, 50 µl viruses were injected into thoracic cavity of 10 days old pups via insulin syringe, avoiding the heart and lungs. For EdU injection, EdU (Sigma, 900584) was dissolved in saline and 5 mg/kg EdU was delivered to mice by intraperitoneal injection for two weeks after AAV transduction. For heart harvesting, mouse was anesthetized by 2% isoflurane and the heart was exposed by opening chest. After that, 15% KCl was injected into inferior vena cava to achieve asystole at diastole, then the heart was rapidly isolated and flushed with D-PBS through LV to wash out blood. Half of the apex was isolated and immersed in RNALater (Qiagen, 76104) at room temperature for RNA extraction, while the other half was snap frozen in liquid nitrogen for protein extraction. The rest part of heart was fixed in 4% paraformaldehyde for 24 hours and subsequently embedded by paraffin.
Liao D., Chen W., Tan C.Y., Wong J.X., Chan P.S., Tan L.W., Foo R, & Jiang J. (2019). Upregulation of Yy1 Suppresses Dilated Cardiomyopathy caused by Ttn insufficiency. Scientific Reports, 9, 16330.
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3

Temporal Analysis of Cell Proliferation

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Cells were plated in six identical plates and allowed to adhere overnight. The next morning all plates were treated identically with inhibitors, and one plate was incubated with EdU (Sigma) for 12 h at a concentration of 10 μM. At the 12-hour mark, this plate was fixed, and EdU was added to a second plate. Every 12 h a subsequent plate was fixed and another was pulsed with EdU to capture the next timepoint. Cell fixation and permeabilization were carried out as described above (Image Cytometry of Cancer Cell Lines). Wells were then treated with a reaction buffer containing 2 mM CuSO4 (Sigma), 8 μM AlexaFluor Azide 647 (LifeTech), and 100 mM sodium ascorbate (Sigma) in PBS and incubated 1 h. After washing with PBS, cells were stained for K19, K14, and VIM, imaged, and analyzed as outlined above (Image Cytometry of Cancer Cell Lines) using nuclear detection of 647-channel signal to quantify cellular EdU levels.
Risom T., Langer E.M., Chapman M.P., Rantala J., Fields A.J., Boniface C., Alvarez M.J., Kendsersky N.D., Pelz C.R., Johnson-Camacho K., Dobrolecki L.E., Chin K., Aswani A.J., Wang N.J., Califano A., Lewis M.T., Tomlin C.J., Spellman P.T., Adey A., Gray J.W, & Sears R.C. (2018). Differentiation-state plasticity is a targetable resistance mechanism in basal-like breast cancer. Nature Communications, 9, 3815.
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4

In vivo Proliferation Assay

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For analysis of proliferation in vivo, mice were pulse labelled with EdU (5-ethynyl-2-deoxyuridine; Sigma-Aldrich) and/or BrdU (5-bromo-2-deoxyuridine; Sigma-Aldrich) at a concentration of 20.53 mg/g or 25 mg/g body weight, respectively. For the induction of Cre activity in Math1-creER T2 mice, pregnant dams or pubs were injected with 1 mg tamoxifen dissolved in corn oil intraperitoneally.
Merk D.J., Ohli J., Merk N.D., Thatikonda V., Morrissy S., Schoof M., Schmid S.N., Harrison L., Filser S., Ahlfeld J., Erkek S., Raithatha K., Andreska T., Weißhaar M., Launspach M., Neumann J.E., Shakarami M., Plenker D., Marra M.A., Li Y., Mungall A.J., Moore R.A., Ma Y., Jones S.J.M., Lutz B., Ertl-Wagner B., Rossi A., Wagener R., Siebert R., Jung A., Eberhart C.G., Lach B., Sendtner M., Pfister S.M., Taylor M.D., Chavez L., Kool M, & Schüller U. (2018). Opposing Effects of CREBBP Mutations Govern the Phenotype of Rubinstein-Taybi Syndrome and Adult SHH Medulloblastoma. Developmental cell, 44(6).
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5

EdU Detection in Organoid Cultures

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For EdU detection, EdU (5 μM, Sigma, St. Louis, MO, USA) was added and incubation was conducted for 0.5 h before harvesting the organoids. The organoids were fixed with 4% PFA for 1 h and allowed to precipitate in 30% sucrose solution for 48 h. The organoids were cryosectioned at 20 µm and incubated with PBS containing 0.3% Triton X-100 at room temperature (RT) for 15 min; these samples were then incubated with Click reaction solution at RT for 30 min, protected from light. After three washes with PBS, these samples were stained with DAPI. The numbers of EdU-positive cells were analyzed by ImageJ.
Zhang R., Lu J., Pei G, & Huang S. (2023). Galangin Rescues Alzheimer’s Amyloid-β Induced Mitophagy and Brain Organoid Growth Impairment. International Journal of Molecular Sciences, 24(4), 3398.
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6

Cell Proliferation and Metabolism Analysis

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Flow cytometry analysis was performed on a Gallios flow cytometer (Beckman Coulter). For cell proliferation assessment, the cells were incubated with 10 μM EdU (Sigma-Aldrich) for 2 hr, and EdU was chemically conjugated to 50 μM Auto 488 (Sigma-Aldrich) for 1 hr and incubated with 2.5 μg/mL DAPI for another 30 min. For glucose uptake assays in cells, cells were grown under normal conditions, and 100 μM 2-NBDG (Invitrogen) was added to the medium for 1 hr.
For mitochondrial mass measurement, MitoTracker Green (Life Technologies) was used to assess the mitochondrial quantity. The cells were incubated for 30 min at 37 C with 50 nM MitoTracker Green before analysis.
Xing F., Luan Y., Cai J., Wu S., Mai J., Gu J., Zhang H., Li K., Lin Y., Xiao X., Liang J., Li Y., Chen W., Tan Y., Sheng L., Lu B., Lu W., Gao M., Qiu P., Su X., Yin W., Hu J., Chen Z., Sai K., Wang J., Chen F., Chen Y., Zhu S., Liu D., Cheng S., Xie Z., Zhu W, & Yan G. (2017). The Anti-Warburg Effect Elicited by the cAMP-PGC1α Pathway Drives Differentiation of Glioblastoma Cells into Astrocytes. Cell reports, 18(2), 468-481.
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7

Quantifying Cell Proliferation in NSCs

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For EdU detection, human NSCs (5000 cells/well) were plated on Matrigel‐coated 96 well plates and cultured for 24 hours. The culture medium was then replaced with the medium containing required chemical for up to 24 hours. EdU (2.5 μM, Sigma‐Aldrich) was added, and cells were incubated for another 0.5 hours. During this period, all the cells entering S‐phase incorporated the thymidine analog. Cells were fixed in 4% paraformaldehyde and stained with EdU (Sigma). Nuclei were labeled with DAPI. The numbers of EdU‐positive cells were analyzed using HTS (Cellomics ArrayScan VTI 700).
Wang Q., Dong X., Lu J., Hu T, & Pei G. (2020). Constitutive activity of a G protein‐coupled receptor, DRD1, contributes to human cerebral organoid formation. Stem Cells (Dayton, Ohio), 38(5), 653-665.
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8

Multiparametric Flow Cytometry Assay

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Flow cytometry (FCM) analysis was performed by using a Gallios flow cytometer (Beckman Coulter). For cell proliferation assessment, the cells were incubated with 10 mM EdU (Sigma-Aldrich) for 2 h, and EdU was chemically conjugated to 50 mM Auto 488 (Sigma-Aldrich) for 1 h and incubated with 1 mg/mL DAPI for another 30 min. For neuronal markers analyses, cells were collected after treatment with vehicle or cAMP activators, and then incubated with 1:100 dilution of MAP2 (#FCMAB318PE, Merk Millipore, Temecula, CA, USA) and 1:100 dilution of TUJ1 (β-Tubulin class III Alexa Fluor 488, #560381, BD Pharmingen, San Diego, CA, USA) for 1 h.
Chen Z., Zhong Y., Chen J., Sun S., Liu W., Han Y., Liu X., Guo C., Li D., Hu W., Zhang P., Chen Z., Chen Z., Mou Y., Yan G., Zhu W., Yin W, & Sai K. (2022). Disruption of β-catenin-mediated negative feedback reinforces cAMP-induced neuronal differentiation in glioma stem cells. Cell Death & Disease, 13(5), 493.
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9

EdU Labeling and Click Chemistry Protocol

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Regular drinking water was removed 24hr prior to NMDA injections and replaced with water containing 5-ethynyl-2′-deoxyuridine (EdU; Sigma; 50 mg/100 mL dH2O) and EdU water was replaced every third day. Mice were maintained on EdU water until 4th day after TSA treatment.
Immunolabeled tissue sections were fixed in 4% formaldehyde in PBS for 5 minutes at room temperature, washed for 10 minutes with PBS, permeabilized with 0.01% Triton X-100 in PBS for 1 minute at room temperature, and washed in PBS for 10 minutes. Sections were incubated for 30 minutes at room temperature in 2M Tris, 50 mM CuSO4, Alexa Fluor 568 or 647 Azide (Thermo Fisher Scientific), and 0.5M ascorbic acid in dH2O. Sections were washed with PBS and further processed for immunofluorescence as required.
Palazzo I., Todd L.J., Hoang T.V., Reh T.A., Blackshaw S, & Fischer A.J. (2022). NFkB-signaling promotes glial reactivity and suppresses Müller glia-mediated neuron regeneration in the mammalian retina. Glia, 70(7), 1380-1401.
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10

Cell Proliferation and Metabolism Analysis

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Flow cytometry analysis was performed on a Gallios flow cytometer (Beckman Coulter). For cell proliferation assessment, the cells were incubated with 10 μM EdU (Sigma-Aldrich) for 2 hr, and EdU was chemically conjugated to 50 μM Auto 488 (Sigma-Aldrich) for 1 hr and incubated with 2.5 μg/mL DAPI for another 30 min. For glucose uptake assays in cells, cells were grown under normal conditions, and 100 μM 2-NBDG (Invitrogen) was added to the medium for 1 hr.
For mitochondrial mass measurement, MitoTracker Green (Life Technologies) was used to assess the mitochondrial quantity. The cells were incubated for 30 min at 37 C with 50 nM MitoTracker Green before analysis.
Xing F., Luan Y., Cai J., Wu S., Mai J., Gu J., Zhang H., Li K., Lin Y., Xiao X., Liang J., Li Y., Chen W., Tan Y., Sheng L., Lu B., Lu W., Gao M., Qiu P., Su X., Yin W., Hu J., Chen Z., Sai K., Wang J., Chen F., Chen Y., Zhu S., Liu D., Cheng S., Xie Z., Zhu W, & Yan G. (2017). The Anti-Warburg Effect Elicited by the cAMP-PGC1α Pathway Drives Differentiation of Glioblastoma Cells into Astrocytes. Cell reports, 18(2), 468-481.
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