Immunocytochemical staining was used to visualize the DSBs and distinguish between the cell cycle stages G1 and S/G2. Four hours before fixation, a nucleoside analogue of thymidine, 5-ethynyl-2′-deoxyuridine (EdU; Thermo Fisher Scientific), was added at a final concentration of 10 μM. The cells were fixed in 4% paraformaldehyde and permeabilized in 0.2% Triton-X (both from Sigma-Aldrich, St. Louis, MO, USA). Overnight incubation with primary antibodies against γH2AX (Biolegend, San Diego, CA, USA) and 53BP1 (Santa Cruz Biotechnology, Dallas, TX, USA) was followed by 1 h of incubation with a secondary antibody conjugated with Alexa 555 (Cell Signaling Technology, Danvers, MA, USA). The samples were stained with the Click-iT® EdU Alexa Fluor® 488 Imaging Kit (Thermo Fisher Scientific) to visualize EdU according to the manufacturer’s instructions. Finally, the nuclei were stained with Hoechst dye (BisBenzemide H33342; 1 μg/ml; Sigma-Aldrich).
5 ethynyl 2 deoxyuridine edu
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany, Australia
5-ethynyl-2′-deoxyuridine (EdU) is a modified nucleoside that can be incorporated into DNA during DNA synthesis. It contains a terminal alkyne group that can be detected using a copper-catalyzed click chemistry reaction.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (9 mentions)
5 bromo 2 deoxyuridine brdu, by Merck Group (9 mentions)
Click it edu imaging kit, by Thermo Fisher Scientific (8 mentions)
Click it edu alexa fluor 488 imaging kit, by Thermo Fisher Scientific (7 mentions)
5 ethynyl 2 deoxyuridine (edu), by Thermo Fisher Scientific (6 mentions)
Thymidine, by Merck Group (5 mentions)
Click it edu alexa fluor 555 imaging kit, by Thermo Fisher Scientific (5 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (4 mentions)
Click it edu alexa fluor 488 flow cytometry assay kit, by Thermo Fisher Scientific (4 mentions)
Click it edu alexa fluor imaging kit, by Thermo Fisher Scientific (4 mentions)
Tamoxifen, by Merck Group (4 mentions)
Alexafluor 647 click it edu kit, by Thermo Fisher Scientific (4 mentions)
Alexa fluor 647 azide, by Thermo Fisher Scientific (3 mentions)
Click it edu alexa fluor 647 imaging kit, by Thermo Fisher Scientific (3 mentions)
Formaldehyde, by Merck Group (3 mentions)
Ascorbic acid, by Merck Group (3 mentions)
Click it reaction cocktail, by Thermo Fisher Scientific (3 mentions)
Click it edu flow cytometry assay kit, by Thermo Fisher Scientific (3 mentions)
P8920, by Merck Group (2 mentions)
Rpmi 1640 medium, by Merck Group (2 mentions)
179 protocols using 5 ethynyl 2 deoxyuridine edu
1
Visualizing DNA Double-Strand Breaks in Cell Cycle
2
Monitoring DNA Repair Kinetics via EdU
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were cultured on 24-mm cover slips and were serum starved (0.5% fetal calf serum) for 2 days to accumulate cells in G0 phase (∼98%). After damage induction, cells were directly labeled with EdU for 3 h (wt cells) or 7 h (GG-NER deficient cells) using Ham's F10 supplemented with 0.5% dialyzed fetal calf serum containing 20 μM 5-ethynyl-2΄-deoxyuridine (EdU, ThermoFisher Scientific). 2΄-Deoxy-5-fluorouridine (1 μM) (Floxuridine, Sigma-Aldrich) was added to inhibit the thymidylate synthase to prevent the generation of endogenous thymidine. After Edu labeling, medium was changed to Ham's F10 supplemented with 0.5% dialyzed fetal calf serum containing 10 μM thymidine (Sigma-Aldrich) for 15 min to deplete unincorporated EdU in the cell. Cells were fixed by incubation with 3.6% formaldehyde (Sigma-Aldrich) in PBS with 0.5% Triton X-100 (Sigma-Aldrich) for 15 min at room temperature. After permeabilization in 0.5% Triton for 20 min, cells were blocked with 3% BSA (Sigma-Aldrich) in PBS.
3
Quantification of Endothelial Cell Proliferation after Injury
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To assess EC proliferation after IR, 5-ethynyl-2′-deoxyuridine (EdU) (Thermo Fisher Scientific, cat. #E10415) was diluted 2.5 mg/mL in sterile PBS and injected intraperitoneally on days 3 and 4 post-injury at a dose of 10 μl/ g body weight. Hearts were harvested from sacrificed animals on day 5 post-injury. Cryosectioning and antibody staining for Erg was performed as described above. To detect endothelial proliferation, the protocol for Click-iT EdU Imaging (Thermo Fisher Scientific, cat. #C10086) was carried out according to manufacturer’s instructions.
4
Demyelination and Remyelination in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Male rats animals 10–12 weeks were anaesthetized with isoflurane and subjected to a stereotactic injection into the corpus callosum (AP: 1.0 mm; L:1.2; DV: 2.2) (Supplementary Fig. 3a ) with 2.5 μL 0.1% lysolecithin (Sigma, St Louis, MO) in PBS over 5 min using a Hamilton syringe 1710RN (Sigma, St Louis, MO)69 (link),70 (link). Animals were kept for 24 h, 72 h, 10 day or 15 day and evaluated with flow cytometry, qPCR, TEM or immunohistochemistry. For in vivo differentiation, 5-ethynyl-2′-deoxyuridine (EdU; Thermo Fisher Scientific, Waltham, MA) was either injected i.p. 25 mg/kg, or added to the drinking water (0.2 mg/mL) for a period of 10 day following LPC injection or in naïve animals for the same time period. Injected animals were kept for 6 weeks.
5
Tissue Proliferation Assessment via EdU Labeling
6
EdU Labeling of Proliferating Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were exposed to 10 µM of 5-ethynyl-2′-deoxyuridine (EdU; Thermo Fisher Scientific) for 1 h at 37 °C. The cells were fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, and reacted with Click Additive Solution (Beyotime, Shanghai, China) for 30 min in the dark. Nuclei were stained with Hoechst 33,342 (Beyotime). Proliferating cells were visualized under a fluorescence microscope.
7
T Cell Proliferation Assay for PBMC Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human PBMCs from five healthy donors were thawed from ImmunXperts biobank and incubated with respective PE/PPE candidates (10µg/mL) for 7 days at 37°C. PBMCs from same donors were incubated with CEF (Mabtech, 3616-1) (HLA class I control) and CEFTA (Mabtech, 3617-1) (HLA class II control) peptide pools, a positive control for T cell activation. On day 6 of culture, the cells were incubated with 5-ethynyl-2’-deoxyuridine (EdU) (Thermofisher) (1μM) for 16h for assessment of T cell proliferation. The day after, supernatants from human PBMC stimulation assays were collected and stored at−80°C for further cytokine analysis. Each cell culture included a set of untreated control wells. Cells were stained for flow cytometry analysis of T cell surface markers (CD4 and CD8), fixed/permeabilized, and the incorporated EdU was stained with a fluorescent azide to assess T cell proliferation by measuring EdU uptake using flow cytometry.
8
Crayfish Hemocyte and HPT Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Five crayfish were injected with 5-ethynyl-2′-deoxyuridine (EdU) (ThermoFisher) at a dose of 15 μg/g body weight. Three days post-injection, 150 μL of hemolymph was taken from each animal and fixed (0 hpw). Five animals were bled as described in 2.3. One hour later, 150 μL of hemolymph was taken from each animal and fixed (1 hpw). Then, the HPT was removed, HPT cells were dissociated by collagenase treatment [4] (link), and fixed with 4% PFA. After fixation for 10 min at room temperature, the cell concentration in each hemolymph sample was measured. THC was determined for each animal as described in 2.3. For EdU staining, fixed cells were collected by centrifugation at 300 g for 3 min, and permeabilized with 0.5% Triton X-100 for 20 min. Cells were then stained with Click-iT™ EdU Alexa Fluor™ 488 Flow Cytometry Assay Kit (ThermoFisher) following the manufacture's instruction. The GC percentage and EdU-positive ratio of circulating hemocytes, and the EdU-positive ratio of HPT cells were analyzed by flow cytometry. For each sample, 10000 cells were analyzed. For all crayfish, EdU was injected into the abdominal cavity, and hemolymph was taken near the site of injection.
9
U937-PR9 Cell Culture and EdU Labeling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U937-PR9 cells were cultured in RPMI-1640 medium (Sigma Aldrich R7388) supplemented with 1% penicillin/streptomycin (Sigma-Aldrich P4333) and 10% fetal bovine serum (Sigma-Aldrich F9665) and maintained at 37 °C and 5% CO2. U937-PR9 were seeded on poly-l -lysine (Sigma-Aldrich P8920) coated glass coverslips immediately before experiments. Cells were incubated with 10 µM of the synthetic nucleoside 5-Ethynyl-2′-deoxyuridine (EdU) (Thermo Fisher Scientific) for 25 min at 37 °C and 5% CO2.
10
Evaluating Neurogenesis and Metabolomics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Supplemental Appendix describes an estimate of motor thresholds for each rTMS intensity, 5-ethynyl-2′-deoxyuridine (EdU; Thermo Fisher Scientific) labeling of newly born cells, behavioral analysis, and brain and serum collection. The Supplemental Appendix also summarizes serotonin and BDNF enzyme-linked immunosorbent assays, analysis of neurogenesis, and metabolomics.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!