EdU (5-ethynyl-2’-deoxyuridine) can be used as a replacement for BrdU (5-bromo-2’-deoxyuridine) and directly measures de novo DNA synthesis or S-phase synthesis of the cell cycle using click chemistry. Click chemistry is a method of covalently coupling an azide with an alkyne. Detection of EdU employs the copper(I) catalyzed click reaction with an azide modified fluorescent dye to form a stable triazole ring. Because of the small size of the click detection reagent, no harsh denaturation steps are needed to gain access to the DNA. Eliminating this step allows for more reproducible results, a simpler and quicker protocol and measurements which can easily be multiplexed with relevant antibody based targets including phospho-histone H3, Ki-67, and cyclin B1 as well as dual-pulse experiments with BrdU by flow cytometry, fluorescence microscopy, microplate (HTS) or high-throughput imaging (HCS).
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
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![Proliferation of ...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/g004686.jpg","Proliferation of muntjac cells detected with Click-iT™ EdU.","Muntjac cells were treated with 10 µM EdU (Cat. No. A10044) for 45 min; cells were then fixed and permeabilized, and EdU that had been incorporated into newly synthesized DNA was detected using the far red–fluorescent Click-iT™ EdU Alexa Fluor® 647 High-Throughput (HCS) Assay (Cat. No. A10208), utilizing the technical tip for converting the HCS assay to conventional fluorescence microscopy. Tubulin was labeled with an anti-α-tubulin antibody (Cat. No. A11126) and visualized with Alexa Fluor® 350 Goat Anti–Mouse IgG (Cat. No. A11045, A21049). The Golgi complex was stained with green-fluorescent Alexa Fluor® 488 conjugate of lectin HPA from <i>Helix pomatia</i> (edible snail) (Cat. No. L11271), and peroxisomes were labeled with an anti-peroxisome antibody and visualized with orange-fluorescent Alexa Fluor® 555 Donkey Anti–Rabbit IgG (Cat. No. A31572).");)
![Proliferating cel...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/Rat%20ileum%20EdU.jpg","Proliferating cells in rat ileum using Click-iT® EdU.","Cell proliferation in a rat ileum tissue section was detected with the Click-iT® EdU Alexa Fluor® 594 Imaging Assay (C10339). Proliferating cells fluoresce red; nuclei are stained with the blue-fluorescent counterstin, Hoechst 33342 (Cat. no. H1399).");)
![Proliferating cel...](javascript: toggle("http://tools.invitrogen.com/content/sfs/gallery/high/EdU%20intestine.JPG","Proliferating cells labeled in vivo with the Click-iT® EdU assay.","EdU was administered to mice intraperitoneally 2 hours before sacrifice. After intestinal tissues were formalin fixed and paraffin embedded, EdU was labeled with the Click-iT® reaction from the Click-iT® EdU Alexa Fluor® 488 Imaging Assay (Cat. no. C10337), using approximately 250 µL of reaction cocktail per slide. The tissue sections were then washed and treated with mounting media containing DAPI, EdU-positive cells are labeled green, nuclei are stained blue and red autofluorescence was enhanced for image contrast.");)
