Click it edu proliferation assay kit

Manufactured by Thermo Fisher Scientific

The Click-iT Edu Proliferation Assay Kit is a tool used to measure cellular proliferation. It utilizes a modified nucleoside that is incorporated into newly synthesized DNA, allowing for the quantification of cell division.

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Lab products found in correlation

Cell counting kit 8 kit, by Beyotime (1 mentions) Accuri c6, by BD (1 mentions) Pi staining buffer, by Merck Group (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions)

Close spelling variants of this product

Click-iT EdU (80 citations) EdU assay kit (30 citations) EdU assay (10 citations)

5 protocols using click it edu proliferation assay kit

Proliferation Assay of HUVECs with MDA-MB-231-HEVs

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The Click-iT™ EdU Proliferation Assay Kit (Invitrogen; Thermo Fisher Scientific, Cat No. C10499) was used for the proliferation assay of HUVECs following the manufacturer’s instruction. Briefly, HUVECs were seeded in 24 wells, plated and incubated overnight. The adhered HUVECs were starved in EBM-2 for 3 h, followed by incubation with 10 µM EdU in either basal EBM-2 or 50 µg/mL MDA-MB-231-HEVs. After 24 h, the HUVECs were fixed by Click-iT EdU fixative. Then, the cell nucleus was stained by Hoechst 33342. By fluorescent imaging, the total cells were counted with nucleus fluorescence and the proliferative cells were counted with EdU fluorescence.

Zhang P., Lim S.B., Jiang K., Chew T.W., Low B.C, & Lim C.T. (2021). Distinct mRNAs in Cancer Extracellular Vesicles Activate Angiogenesis and Alter Transcriptome of Vascular Endothelial Cells. Cancers, 13(9), 2009.

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Cell Proliferation Monitoring Assay

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Cell proliferation was monitored using a Click-iT Edu Proliferation assay kit (#C10499, Invitrogen). About 10,000 cells/well were plated in a 96-well plate the day before the experiment, and the manufacturing instructions were followed.

Potenza F., Cufaro M.C., Di Biase L., Panella V., Di Campli A., Ruggieri A.G., Dufrusine B., Restelli E., Pietrangelo L., Protasi F., Pieragostino D., De Laurenzi V., Federici L., Chiesa R, & Sallese M. (2021). Proteomic Analysis of Marinesco–Sjogren Syndrome Fibroblasts Indicates Pro-Survival Metabolic Adaptation to SIL1 Loss. International Journal of Molecular Sciences, 22(22), 12449.

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Measuring Cell Proliferation Dynamics

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Cell proliferation was measured by CCK-8 assay and EdU incorporation assay. For CCK-8 assay, A172 and U251 cells were seeded in 96-well plates, and cell viability was eveluated using the Cell Counting Kit-8 kit (C0038, Beyotime) according to the manufacturer’s instructions. Cell viability was expressed as the percentage of control group. The EdU incorporation assay was conducted as previously reported [18 (link)]. In brief, proliferating A172 and U251 cells cultured in 24-well plates were examined using the Click-iT Edu Proliferation Assay Kit (C10337, ThermoFisher Scientific) according to the manufacturer’s instructions. The Edu incorporation was analyzed by the flow cytometry (Accuri C6, BD). Each group included five replicates and three independent experiments were performed.

Chen W., Hong L., Hou C., Wang Y., Wang F, & Zhang J. (2020). MicroRNA-585 inhibits human glioma cell proliferation by directly targeting MDM2. Cancer Cell International, 20, 469.

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Cell Cycle Analysis of Murine Neurospheres

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Cell cycle analysis was performed using a Click-iT EdU proliferation assay kit (ThermoFisher, C10636), following manufacturer’s instructions. Briefly, primary murine neurospheres with fewer than 8 passages were dissociated with Accutase to generate single cell suspensions. 1 × 10⁵ cells were seeded in a well of a 12-well tissue culture plate and incubated overnight in the neurosphere medium. At the end of the 23rd hr, the medium was completely replaced with a fresh medium containing 10 μM Edu reagent. Cells were immediately returned to the incubator and grown for one more hour. The cells were lifted with Accutase, fixed with 4% PFA, and permeabilized with saponin, before the EdU was conjugated to a fluorophore with catalysis of a copper compound via a “click chemistry” reaction. The cells were then incubated in a PI staining buffer (200 μg/ml, Sigma, P4864) for 30 min at RT and analyzed on a BD LSR II flow cytometer.

Chen Z., Herting C.J., Ross J.L., Gabanic B., Vallcorba M.P., Szulzewsky F., Wojciechowicz M.L., Cimino P.J., Ezhilarasan R., Sulman E.P., Ying M., Ma’ayan A., Read R.D, & Hambardzumyan D. (2020). Genetic driver mutations introduced in identical cell-of-origin in murine glioblastoma reveal distinct immune landscapes but similar response to checkpoint blockade. Glia, 68(10), 2148-2166.

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EdU Proliferation Assay of HUVECs

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The Click-iT™ EdU Proliferation Assay kit (Thermo Fisher Scientific, Cat No. C10499) was used for the proliferation assay of HUVECs following the manufacturer's instruction. Briefly, HUVECs were seeded in 24 well plated and incubated for overnight. The adhered HUVECs were starved in EBM-2 for 3 hours, followed by incubation with 10 µM EdU in either basal EBM-2 or 50 µg/mL MDA-MB-231-HEXOs. After 24 hours, the HUVECs were fixed by Click-iT EdU fixative. Then the cell nucleus was stained by Hoechst 33342. By fluorescent imaging, the total cells were counted with nucleus fluorescence and the proliferative cells were counted with EdU fluorescence.

Zhang P., Lim S.B., Jiang K., Chew T.W., Low B.C., & Lim C.T. (2020). Cancer exosomes harbor diverse hypoxia-targeted mRNAs and contribute toward tumor angiogenesis.

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