5 ethynyl 2 deoxyuridine edu (Jena Bioscience)

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Name:

5 Ethynyl 2 deoxyuridine 5 EdU

Description:

5 EdU 5 Ethynyl 2 deoxyuridine can be used as a replacement for BrdU 5 Bromo 2 deoxyuridine to measure de novo DNA synthesis during the S phase of the cell cycle 5 EdU is cell permeable and incorporates into replicating DNA instead of its natural analog thymidine The resulting ethynyl functionalized DNA can subsequently be detected via Cu I catalyzed click chemistry that offers the choice to introduce a Biotin group via Azides of Biotin for subsequent purification tasks or a fluorescent group via Azides of fluorescent dyes for subsequent microscopic imaging 1 5 Presolski et al 6 and Hong et al 7 provide a general protocol for Cu I catalyzed click chemistry reactions that may be used as a starting point for the set up and optimization of individual assays

Catalog Number:

clk-n001-100

Molecular Weight:

252.23 g/mol

Price:

152.25

Applications:

DNA synthesis monitoring[1-5]

Purity:

≥ 98 % (HPLC)

Category:

Click Chemistry

Format:

white to off-white solid

Formula:

C11H12N2O5

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Structured Review

Jena Bioscience 5 ethynyl 2 deoxyuridine edu

Nuclear ROCK1 colocalizes with Hsc70 at nucleolus. (A and B) Fibroblasts were infected with the Merlin strain and stained at 72 hpi for ROCK1 (red) and DAPI (blue). Replication compartments were imaged either by metabolically labeling nascent DNA with <t>ethynyl-2′-deoxyuridine</t> <t>(EdU)</t> followed by click-mediated fluorescent labeling of EdU (green) (A) or by staining for UL57 (green) (B). (C) Fibroblasts were mock infected and stained for Hsc70 (green) and DAPI (blue) or infected with the Merlin strain and stained at 72 hpi for ROCK1 (red), Hsc70 (green), pp28 (violet), and DAPI (blue). (D) Fibroblasts were infected with the Merlin strain and stained at 72 hpi for ROCK1 (red), Hsc70 (green), the nucleolar marker fibrillarin (violet), and DAPI (blue). Scale bars are 10 μm.

5 ethynyl 2 deoxyuridine edu - by Bioz Stars, 2020-07

93/100 stars

Images

1) Product Images from "Rho-Associated Coiled-Coil Kinase 1 Translocates to the Nucleus and Inhibits Human Cytomegalovirus Propagation"

Article Title: Rho-Associated Coiled-Coil Kinase 1 Translocates to the Nucleus and Inhibits Human Cytomegalovirus Propagation

Journal: Journal of Virology

doi: 10.1128/JVI.00453-19

Figure Legend Snippet: Nuclear ROCK1 colocalizes with Hsc70 at nucleolus. (A and B) Fibroblasts were infected with the Merlin strain and stained at 72 hpi for ROCK1 (red) and DAPI (blue). Replication compartments were imaged either by metabolically labeling nascent DNA with ethynyl-2′-deoxyuridine (EdU) followed by click-mediated fluorescent labeling of EdU (green) (A) or by staining for UL57 (green) (B). (C) Fibroblasts were mock infected and stained for Hsc70 (green) and DAPI (blue) or infected with the Merlin strain and stained at 72 hpi for ROCK1 (red), Hsc70 (green), pp28 (violet), and DAPI (blue). (D) Fibroblasts were infected with the Merlin strain and stained at 72 hpi for ROCK1 (red), Hsc70 (green), the nucleolar marker fibrillarin (violet), and DAPI (blue). Scale bars are 10 μm.

Techniques Used: Infection, Staining, Metabolic Labelling, Labeling, Marker

2) Product Images from "γH2AX in the S Phase After UV Irradiation Corresponds to the Sites of DNA Replication and Not DNA Damage"

Article Title: γH2AX in the S Phase After UV Irradiation Corresponds to the Sites of DNA Replication and Not DNA Damage

Journal: bioRxiv

doi: 10.1101/810689

$UV-induced γH2AX does not report on the extent of DNA damage in the S phase cells (A) Definition of the colocalization metric: more than 50% overlap between the foci from the two channels at a position. Images are of NCS-treated cells: those 53BP1 foci colocalizing with γH2AX foci are considered as DSBs (B) Percentage of γH2AX foci which are double strand breaks for control, UV- and NCS-treated cells. For UV treatment a very little fraction of γH2AX foci in the S phase cells corresponds to DSBs. (C) Global transcription is measured in UV-treated cells by quantifying 5-ethynyl-uridine incorporation. (D) Bar graphs for γH2AX and EU are normalized across the four dosages. Resultant bar graphs for EU are inverted over those for γH2AX such that the total height of the two bars corresponding to control population in every cell cycle phase is unity. A clear gap is observed between γH2AX and EU bars for G1 and G2/M phases after UV treatment. The γH2AX bars in the S phase not just overlap with EU bars but goes past the unit box showing a disproportionate increase in γH2AX. (E) Mean levels of γH2AX increase with the increase in DNA damage as achieved by increasing the UV dosage in all the phases of the cell cycle. For all the dosages, there is a sharp γH2AX peak in the S phase. (F) Increase in DNA damage always leads to decrease in EU as seen across the population treated with different dosages of UV. But within a population there is no dip in EU levels corresponding to the γH2AX peak showing that γH2AX in the S phase after UV does not reflect on the total extent of DNA damage in those cells. Cells were treated with 10 J/m 2 and 1.6 µg/ml NCS for two minutes. Line graphs are showing relative levels with respect to the mean value of G1 phase in control cells. Scale bar: 10 µm.$
Figure Legend Snippet: UV-induced γH2AX does not report on the extent of DNA damage in the S phase cells (A) Definition of the colocalization metric: more than 50% overlap between the foci from the two channels at a position. Images are of NCS-treated cells: those 53BP1 foci colocalizing with γH2AX foci are considered as DSBs (B) Percentage of γH2AX foci which are double strand breaks for control, UV- and NCS-treated cells. For UV treatment a very little fraction of γH2AX foci in the S phase cells corresponds to DSBs. (C) Global transcription is measured in UV-treated cells by quantifying 5-ethynyl-uridine incorporation. (D) Bar graphs for γH2AX and EU are normalized across the four dosages. Resultant bar graphs for EU are inverted over those for γH2AX such that the total height of the two bars corresponding to control population in every cell cycle phase is unity. A clear gap is observed between γH2AX and EU bars for G1 and G2/M phases after UV treatment. The γH2AX bars in the S phase not just overlap with EU bars but goes past the unit box showing a disproportionate increase in γH2AX. (E) Mean levels of γH2AX increase with the increase in DNA damage as achieved by increasing the UV dosage in all the phases of the cell cycle. For all the dosages, there is a sharp γH2AX peak in the S phase. (F) Increase in DNA damage always leads to decrease in EU as seen across the population treated with different dosages of UV. But within a population there is no dip in EU levels corresponding to the γH2AX peak showing that γH2AX in the S phase after UV does not reflect on the total extent of DNA damage in those cells. Cells were treated with 10 J/m 2 and 1.6 µg/ml NCS for two minutes. Line graphs are showing relative levels with respect to the mean value of G1 phase in control cells. Scale bar: 10 µm.

Techniques Used:

Synthesized:

Article Title: Dual Roles of Poly(dA:dT) Tracts in Replication Initiation and Fork Collapse
Article Snippet: .. OK-seq was performed in asynchronous mouse B cells activated for 48 hours, as previously described in a detailed protocol Here, the exponentially growing B cells were pulsed with 20 mM EdU (5-ethynyl-2’-deoxyuridine, Jenabioscience) for 2 minutes to label newly synthesized DNA. ..

Generated:

Article Title: Truncated PPM1D impairs stem cell response to genotoxic stress and promotes growth of APC-deficient tumors in the mouse colon
Article Snippet: .. Cell cycle checkpoint Cells were pulsed with 5-ethynyl-2′-deoxyuridine (EdU, Jena Bioscience) and subsequently exposed to ionizing radiation generated by Precision X-RAD 225XL (dose 3, 6, or 10 Gy) and were grown for further 20 h in the presence of nocodazole (250 ng/mL). .. Cells were stained with anti-pSer10-histone H3 (mitotic marker, Santa Cruz) and 4′,6-diamidino-2-phenylindole (DAPI as a marker of DNA content) and EdU was labeled with AlexaFluor 647 using Click-iT reaction (Thermo Scientific).

Mouse Assay:

Article Title: Notch3 is necessary for neuronal differentiation and maturation in the adult spinal cord
Article Snippet: .. EdU labelling Two-month-old mice were injected daily for 7 days with 5-ethynyl-2-deoxyuridine (EdU, 4 mg/kg i.p., Jena Bioscience, Jena, Germany). .. On the 8th day, the mice were killed by perfusion-fixation with 4% paraformaldehyde/PBS under anaesthesia.

Article Title: Peripheral nerve injury induces adult brain neurogenesis and remodelling
Article Snippet: .. EdU labelling Six‐week‐old mice (n = 5) were subjected to CCI, then 5 weeks later were injected twice daily for 3 days with 5‐Ethynyl‐2‐deoxyuridine (EdU; 4 mg/kg i.p., Jena Bioscience, Jena, Germany) . .. Four days after last injection, the mice were killed by perfusion‐fixation under anaesthesia.

Incubation:

Article Title: Rho-Associated Coiled-Coil Kinase 1 Translocates to the Nucleus and Inhibits Human Cytomegalovirus Propagation
Article Snippet: .. Briefly, cells were incubated with 10 μM 5-ethynyl-2′-deoxyuridine (EdU) (Jena Bioscience GmbH) for 30 min. .. Cells were then fixed with 4% formaldehyde for 10 min, permeabilized with 0.5% Triton X-100 for 20 min, and stained with staining mix (100 mM Tris, pH 8.5, 1 mM CuSO4 , 10 μM fluorescent azide, 100 mM ascorbic acid) for 30 min. EdU-stained cells were immunostained for ROCK1 and DAPI as described above.

Injection:

Recombinant:

Article Title: Immature spinal cord neurons are dynamic regulators of adult nociceptive sensitivity
Article Snippet: .. Reagents Anti-fibroblast growth factor 2 (FGF2) neutralizing antibody, recombinant FGF2, recombinant epidermal growth factor (EGF; R & D Systems, Minneapolis, MN, USA), Erlotinib (ChemieTek LLC, Indianapolis, IN, USA), recombinant brain-derived neurotrophic factor (BDNF) (Prospec Bio, East Brunswick, NJ, USA), 7,8-dihydroxyflavone (TCI America, Philadelphia, PA, USA), 5-Ethynyl-2-deoxyuridine (EdU), azide-fluorescein (Jena Bioscience, Jena, Germany). .. Antibodies: Mash1, doublecortin (DCX), Notch3, nestin, calretinin, TrkB, glial fibrillary acidic protein, Skp2 (Santa Cruz Biotech, Dallas, TX, USA), NeuN (Millipore, Billerica, MA, USA), Cy3- and fluorescein isothiocyanate (FITC)-linked secondary antibodies (Jackson Immuno-Research, West Grove, PA, USA).