Edu proliferation assay kit

Manufactured by Abcam

The EdU Proliferation Assay Kit is a laboratory tool used to measure cell proliferation. It detects the incorporation of the synthetic nucleoside EdU (5-ethynyl-2'-deoxyuridine) into the DNA of dividing cells, enabling the identification and quantification of actively proliferating cells.

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EdU assay kit (8 citations) EdU assay (4 citations) EdU assay/EdU Staining Proliferation Kit (iFluor 488) (2 citations) EdU Assay / EdU Staining Proliferation Kit (2 citations)

2 protocols using edu proliferation assay kit

EdU labeling of chick epiblast

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Embryos, isolated with a ring filter paper support and with a node graft at st. 4, were incubated at 38°C in the orientation of the ventral side upward. After 6 or 8 h, the embryos were flipped to dorsal side facing upward, the vitelline membrane was pierced, 200 µl of 40 µM EdU was added to the epiblast side of the embryos, and the embryos were incubated in this orientation for a further 2 h. Only the epiblast cells were labeled using this procedure. We confirmed that these dorsal-side-upward cultures allowed embryo development at least up to st. 15, similar to ordinary ventral-side-upward cultures. After fixation with 4% PFA for 15 min at room temperature, the embryo specimens were processed for Alexa 488 fluorescence using the EdU Proliferation Assay Kit (Abcam) and counterstained with 1 μg/ml 4′,6-diamidino-2-phenylindole (DAPI). Fluorescence images were taken using an FV3000 inverted laser microscope (Olympus).

Yoshihi K., Kato K., Iida H., Teramoto M., Kawamura A., Watanabe Y., Nunome M., Nakano M., Matsuda Y., Sato Y., Mizuno H., Iwasato T., Ishii Y, & Kondoh H. (2022). Live imaging of avian epiblast and anterior mesendoderm grafting reveals the complexity of cell dynamics during early brain development. Development (Cambridge, England), 149(6), dev199999.

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Quantifying DNA Synthesis via EdU Labeling

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To measure the rate of EdU (5-ethynyl-2ʹ-deoxyuridine) incorporation into DNA, cells were grown on hydrogels for 24 hours, after which they were prepared into fluorescence microscopy samples using an EdU Proliferation Assay Kit (Abcam, ab222421), according to the manufacturer’s instructions. Briefly, the cells were supplemented with 20 μM EdU for 2 hours, fixed, permeabilized, and the EdU was stained with iFluor 647 azide via a copper-catalyzed click reaction. Nuclei were counterstained before imaging (see below).

Isomursu A., Park K.Y., Hou J., Cheng B., Mathieu M., Shamsan G., Fuller B., Kasim J., Mohsen Mahmoodi M., Lu T.J., Genin G.M., Xu F., Lin M., Distefano M., Ivaska J, & Odde D.J. (2022). Negative durotaxis: cell movement toward softer environments. Nature materials, 21(9), 1081-1090.

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