Click it edu alexa fluor 594 imaging kit

Manufactured by Thermo Fisher Scientific

Sourced in United States, United Kingdom

The Click-iT EdU Alexa Fluor 594 Imaging Kit is a fluorescence-based tool for detecting and visualizing DNA synthesis in cells. It provides a sensitive and specific way to measure cell proliferation through the incorporation of the nucleoside analog EdU (5-ethynyl-2'-deoxyuridine) into newly synthesized DNA, which is then detected by the Alexa Fluor 594 fluorescent dye.

Automatically generated - may contain errors

Lab products found in correlation

128 protocols using click it edu alexa fluor 594 imaging kit

Cell Proliferation and Senescence Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

To determine the percentage of cells in S-phase, proliferating, quiescent or senescent A1 cells were sub-cultured and incubated in growth medium supplemented with 5 µM 5-ethynyl-2′-deoxyuridine (EdU) for 24 hr. Cells were then fixed in glutaraldehyde and SA-β-gal staining performed as described. Next, cells were fixed in 4%PFA for 10 min and stained using Click-iT Edu Alexa Fluor 594 Imaging kit (Life Technologies, UK) according to manufacturer's instructions.
To detect EdU incorporation in salamander tissues, 10 mM EdU (20 µl per animal) were administered to regenerating newts by i.p. injection. After 48 hr, blastemas were collected, cryosectioned and stained to determine SA-β-gal activity as described. Sections were fixed in 4% PFA for 10 min and EdU incorporation determined using Click-iT Edu Alexa Fluor 594 Imaging kit (Life Technologies).

Yun M.H., Davaapil H, & Brockes J.P. (2015). Recurrent turnover of senescent cells during regeneration of a complex structure. eLife, 4, e05505.

+ Open protocol

+ Expand

Evaluating Cell Growth Inhibition

Check if the same lab product or an alternative is used in the 5 most similar protocols

To test the growth activity of U87MG and LN229 cells, Click-iT EdU Alexa Fluor 594 Imaging Kit (Thermo Fisher Scientific, Massachusetts, USA) was used according to the manufacturer’s protocol. After treated with 5 μM GSK343 or 0.1% DMSO for 48 h, the proportion of cells incorporated EdU was determined with fluorescence microscopy (Nikon, Tokyo, Japan).

Han D., Yu T., Dong N., Wang B., Sun F, & Jiang D. (2019). Napabucasin, a novel STAT3 inhibitor suppresses proliferation, invasion and stemness of glioblastoma cells. Journal of Experimental & Clinical Cancer Research : CR, 38, 289.

+ Open protocol

+ Expand

Induction of Cell Cycle Arrest and DNA Damage

Check if the same lab product or an alternative is used in the 5 most similar protocols

After 18 dye + light treatments or 16 h treatment with 0.4 μM APH, cells were allowed to recover for 8h prior to an 8h incubation with 7 μM Cdk1 inhibitor RO3306 (Millipore). Where indicated, cells were incubated with MRE11 inhibitor Mirin (100 μM; Sigma) or RAD51 inhibitor B02 (100 μM; Sigma) during the last hour of Cdk1 inhibitor incubation. Cells were washed twice with PBS and incubated in fresh media containing 1× EdU and 50ng/ml concentrations colcemid with or without MRE11 or RAD51 inhibitor for 1h prior to harvest. Metaphase spreads were prepared as described above and EdU staining performed using Click-iT™ EdU Alexa Fluor™ 594 imaging kit (ThermoFisher) after telomere FISH staining.

Fouquerel E., Barnes R.P., Uttam S., Watkins S.C., Bruchez M.P, & Opresko P.L. (2019). Targeted and Persistent 8-Oxoguanine Base Damage at Telomeres Promotes Telomere Loss and Crisis. Molecular cell, 75(1), 117-130.e6.

+ Open protocol

+ Expand

S100A11 Regulates Cell Proliferation

Check if the same lab product or an alternative is used in the 5 most similar protocols

DNA synthesis was monitored by the incorporation of EdU into DNA. Cells were inoculated at a density of 2 × 10⁵ cells per well into six-well plates and cultured in D/F medium with 10% FBS for 24 h and then starved with serum-free D/F medium for an additional 24 h. The starved cell cultures were treated with low-serum D/F medium (0.5% FBS) containing S100A11 (final concentration, 10 μg/ml) for 6 h, 12 h, and 24 h. EdU was added to the cultures 1 h before fixation of the cells, and the incorporated EdU was stained using a Click-iT™ EdU Alexa Fluor™ 594 Imaging Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS; Promega Biosciences) was used for assessment of cell proliferation. Cells were treated with S100A11 by the same method as that described above for DNA synthesis except for differences in the number of cells (1 × 10⁴ cells) and culture plates (96-well plates).

Takamatsu H., Yamamoto K.I., Tomonobu N., Murata H., Inoue Y., Yamauchi A., Sumardika I.W., Chen Y., Kinoshita R., Yamamura M., Fujiwara H., Mitsui Y., Araki K., Futami J., Saito K., Iioka H., Ruma I.M., Putranto E.W., Nishibori M., Kondo E., Yamamoto Y., Toyooka S, & Sakaguchi M. (2019). Extracellular S100A11 Plays a Critical Role in Spread of the Fibroblast Population in Pancreatic Cancers. Oncology Research, 27(6), 713-727.

+ Open protocol

+ Expand

Glioma Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

To assess the proliferative activity of U87 and LN229 glioma cells, Click-iT EdU Alexa Fluor 594 Imaging Kit (Thermo Fisher Scientific, Massachusetts, USA; Cat. No. C10339) was used according to the manufacturer's protocol. After treated with 5 μM GSK343 or 0.1% DMSO for 48h, the proportion of glioma cells incorporated EdU was determined with fluorescence microscopy (Nikon, Tokyo, Japan) at x200 magnification.

Yu T., Wang Y., Hu Q., Wu W., Wu Y., Wei W., Han D., You Y., Lin N, & Liu N. (2017). The EZH2 inhibitor GSK343 suppresses cancer stem-like phenotypes and reverses mesenchymal transition in glioma cells. Oncotarget, 8(58), 98348-98359.

+ Open protocol

+ Expand

EdU Labeling Assay in Drosophila

Check if the same lab product or an alternative is used in the 5 most similar protocols

The EdU labeling assay was modified from previously described with a Click-iT EdU Alexa Fluor 594 Imaging Kit (Thermo Fisher Scientific, C10339) [58 (link)]. Male flies were fed with 100 μM EdU and 5% sucrose for 24 hr. The EdU incorporation in gut was visualized by Click reaction according to the manufacturer’s instruction.

Xi J., Cai J., Cheng Y., Fu Y., Wei W., Zhang Z., Zhuang Z., Hao Y., Lilly M.A, & Wei Y. (2019). The TORC1 inhibitor Nprl2 protects age-related digestive function in Drosophila. Aging (Albany NY), 11(21), 9811-9828.

+ Open protocol

+ Expand

Enteroid Proliferation Assay with IL-22

Check if the same lab product or an alternative is used in the 5 most similar protocols

Enteroids in Matrigel were stimulated with interleukin-22 (10 ng/mL) for 24 hours. EdU (Click-iT EdU Alexa Fluor 594 Imaging Kit, Thermo Fisher C10339) was added at 10 μM 2 hours before fixation. EdU staining was performed as indicated by the manufacturer.

Wong J., Garcia-Carbonell R., Zelic M., Ho S.B., Boland B.S., Yao S.J., Desai S.A., Das S., Planell N., Harris P.A., Font-Burgada J., Taniguchi K., Bertin J., Salas A., Pasparakis M., Gough P.J., Kelliher M., Karin M, & Guma M. (2019). RIPK1 Mediates TNF-Induced Intestinal Crypt Apoptosis During Chronic NF-κB Activation. Cellular and Molecular Gastroenterology and Hepatology, 9(2), 295-312.

+ Open protocol

+ Expand

Quantifying Muscle Stem Cell Proliferation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were injected with EdU (100mg/kg) IP, following the experimental design shown in Figure 3. MuSCs were plated overnight following sorting on laminin-coated 8-well chamber slides (~2500 cells) in DMEM/F12 supplemented with 15% FBS, 1X antibiotic/antimycotic, 1X Glutamax, 1X NEAA. Cells were fixed the following day in 3.7% formaldehyde/PBS for 15min and processed for EdU incorporation using the Click-iT EdU Alexa Fluor 594 Imaging kit (ThermoFisher), according to the manufacturer’s instructions.

Tichy E.D., Ma N., Sidibe D., Loro E., Kocan J., Chen D.Z., Khurana T.S., Hasty P, & Mourkioti F. (2021). Persistent NF-κB activation in muscle stem cells induces proliferation-independent telomere shortening. Cell reports, 35(6), 109098.

+ Open protocol

+ Expand

Multimodal Analysis of Hair Matrix

Check if the same lab product or an alternative is used in the 5 most similar protocols

Slides with comparable tissue sections of the human hair matrix were selected for staining and allowed to air dry at room temperature for 10 min. Slides were subsequently fixed in ice-cold acetone for 10 min and left to air dry for a further 10 min. Each section was circumscribed using a hydrophobic barrier PAP pen (#H4000 Vector Laboratories).
Incorporated EdU/EU was detected as per kit instructions (Click‐iT™ EdU Alexa Fluor™ 594 Imaging Kit, #C10339, Thermo Fisher Scientific; Click‐iT™ RNA Alexa Fluor™ 594 Imaging Kit, #C10330, Thermo Fisher Scientific).
Primary antibodies were diluted in PBS (see below) and were applied to sections; slides were then incubated at 4 °C overnight. Slides were then washed three times in PBS for 2 min and secondary fluorescent antibody was applied at 1:200 in PBS for 45 min at room temperature (anti-mouse/rabbit Alexa Fluor 488/594). Slides were then washed three times in PBS for 2 min and treated with Hoechst 33342 (1:1000 in PBS) (#H3570, Thermo Fisher Scientific). Slides were finally washed in PBS for 2 min and mounted using aqueous mounting medium (#S3025, Dako).

Mitchell E., Mellor C.E, & Purba T.S. (2020). XMU-MP-1 induces growth arrest in a model human mini-organ and antagonises cell cycle-dependent paclitaxel cytotoxicity. Cell Division, 15, 11.

+ Open protocol

+ Expand

Optic Glioma Cell Proliferation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ten thousand optic glioma cells were seeded to fibronectin (10 μg ml⁻¹)-coated 8-chamber slides (Thermo Fisher Scientific, 154534PK), and grown in NSC medium without N2, FGF and EGF for 48 h, supplemented with vehicle (25 mM Tris-HCl, pH 7.3, 100 mM glycine and 10% glycerol), NLGN3 (70 nM unless otherwise noted, OriGene TP307955), or BDNF (70 nM, PeproTech 450-02). EdU proliferation assay was performed using the Click-iT EdU Alexa Fluor 594 imaging kit (Thermo Fisher Scientific C10339) according to the manufacturer’s instructions. EdU was added 8 h before fixing the cells in 4% paraformaldehyde. Images were taken using a Zeiss Axio Imager M2 and the Stereo Investigator software (mbfbioscience v2019). Cell proliferation was determined by dividing the number of EdU⁺ cells by the number of DAPI⁺ cells using Cell Profiler (v.3.1.9). All experiments were repeated three times with similar results.

Pan Y., Hysinger J.D., Barron T., Schindler N.F., Cobb O., Guo X., Yalçın B., Anastasaki C., Mulinyawe S.B., Ponnuswami A., Scheaffer S., Ma Y., Chang K.C., Xia X., Toonen J.A., Lennon J.J., Gibson E.M., Huguenard J.R., Liau L.M., Goldberg J.L., Monje M, & Gutmann D.H. (2021). NF1 mutation drives neuronal activity-dependent initiation of optic glioma. Nature, 594(7862), 277-282.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!