In Vivo EdU Cell Proliferation Assay for High-throughput Screening

Product Information

Accurate quantification of DNA synthesis in vivo for high-throughput applications

The In Vivo ClickTech EdU Cell Proliferation Kit for High-Throughput Screening (HTS) enables precise detection of cell proliferation in whole organisms, supporting studies on tumor growth, neural and tissue development, and cellular regeneration. The assay uses the thymidine analogue EdU (5-ethynyl-2′-deoxyuridine), incorporated into newly synthesized DNA during the S-phase of the cell cycle. Detection is achieved through bioorthogonal ‘click’ chemistry, which fluorescently tags EdU-labeled DNA without denaturation. This ensures bright, stable signals with minimal background, ideal for high-content analysis across diverse tissue types and organisms.

How this kit differs from standard HTS kits

Unlike standard EdU HTS kits designed for cultured cells, the in vivo kit provides increased EdU quantities for systemic administration and includes protocols optimized for animal and tissue samples. Detection reagents are tailored for complex biological matrices, ensuring reproducibility and compatibility with imaging workflows.

How does the ClickTech EdU Cell Proliferation Assay work?

5-ethynyl-2′-deoxyuridine (5-EdU) is a cell-permeable derivative of thymidine in which the methyl group has been replaced by an ethynyl group. EdU is converted to the triphosphate 5-EdUTP and incorporated into newly synthesized DNA during replication. Bioorthogonal copper-catalyzed azide–alkyne cycloaddition (CuAAC) ‘click’ chemistry ^{[1, 2]} is then used to covalently link azide-modified fluorophores to the EdU for detection. This produces highly specific nuclear labeling of proliferating cells with excellent spatial resolution.^[3]

Why choose ClickTech In Vivo HTS?

• Exceptional sensitivity with low background

Click chemistry-based dye conjugation ensures a high signal-to-noise ratio; no cross-reactivity to other biomolecules present in a tissue.

• Faster, simpler workflow

No DNA denaturation or antibody steps necessary.

• No harsh treatments or specialized equipment needed

Tissue morphology and antigenicity are preserved — ideal for multiplexing with immunostaining.

What are the features & applications of the in vivo ClickTech EdU HTS assay?

• Suitable for imaging in mice, rats, zebrafish, and more
• Compatible with injection, water-based delivery or incubation
• Antibody-free, fast detection (~30 minutes)
• Works with multiplex labeling and co-staining

Which organisms can the In Vivo ClickTech EdU Cell Proliferation Assay be used on?

This assay has been used with many different types of organisms, ranging from prokaryotic to eukaryotic. It can be used with animals such as mice, rats, C. elegans and zebrafish.^[4]

LITERATURE

1. Peptidotriazoles on Solid Phase: [1,2,3]-Triazoles by Regiospecific Copper(I)-Catalyzed 1,3-Dipolar Cycloadditions of Terminal Alkynes to Azides. C. W. Tornøe, C. Christensen, M. Meldal, 2002, J. Org. Chem., Vol. 67, p. 3057–3064.
2. A Stepwise Huisgen Cycloaddition Process: Copper(I)-Catalyzed Regioselective “Ligation” of Azides and Terminal Alkynes. V. V. Rostovtsev, L. G. Green, V. V. Fokin, K. B. Sharpless, 2002, Angew. Chemie Int. Ed., Vol. 41, p. 2596–2599.
3. A novel multicolor immunostaining method using ethynyl deoxyuridine for analysis of in situ immunoproliferative response, Y. Kitazawa, H. Ueta, T. Hünig, Y. Sawanobori, K. Matsuno, 2015, Cell Biol., Vol. 144, p. 195–208.
4. Skin cells undergo asynthetic fission to expand body surfaces in zebrafish, K. Y. Chan, C.-C. Sanders Yan, H.-Y. Roan, S.-C. Hsu, T.-L. Tseng, C.-D. Hsiao, C.-P. Hsu & C.-H. Chen, 2022, Nature, Vol. 605, p. 119–125.
5. An efficient protocol for in vivo labeling of proliferating epithelial cells, C. Michel et al., 2018, J. Immunol. Methods, Vol. 457, p. 82-86. https://doi.org/10.1016/j.jim.2018.03.015
6. Cisplatin-induced DNA double-strand breaks promote meiotic chromosome synapsis in PRDM9-controlled mouse hybrid sterility, L. Wang et al., 2018, eLife, Vol. 7, p. e42511. https://doi.org/10.7554/eLife.42511
7. FXR Regulates Intestinal Cancer Stem Cell Proliferation, T. Fu et al., 2019, Cell, Vol. 176, p. 1098-1112. https://doi.org/10.1016/j.cell.2019.01.036