Click chemistry is a ground-breaking chemical method used to attach a substrate of interest to a biomolecule with unprecedented specificity. The creators of this technique (Sharpless, Meldal, and Bertozi) were recently awarded the 2022 Nobel Prize in Chemistry as it already has had a significant impact on the research community in several application areas.
Thermo Fisher Scientific utilizes this Nobel-winning technology in two unique ways, first to transform cell proliferation assay capabilities and second, to offer a highly effective solution for site-specific antibody labelling.
Why is Click Chemistry useful to researchers?
Drug discovery and disease models, particularly for oncology research, are better suited to assess cell proliferation in model organisms rather than dissociated cell culture. However, due to the ubiquitous distribution of amines and thiols within cells and their presence in cell culture, in situ labeling of proteins and newly synthesized nucleic acids is often difficult. Click chemistry labeling technology overcomes this obstacle by employing bioorthogonal reaction partners that are not endogenous in cells, tissues, or model organisms to allow in vivo detection of proliferating cells.
How can Click Chemistry help measure cell proliferation for animal models?
Utilizing this innovative technology, Thermo Fisher Scientific features Click Chemistry in our Click-iT EdU (5-ethynyl-2’-deoxyuridine) and Click-iT Plus EdU Cell Proliferation Kits. These kits offer all the tools necessary for assessing in vivo proliferation and fluorescent EdU detection with a wide range of fluorophores to help meet the needs of researchers. When click chemistry is used in a cell proliferation assay such as Click-iT EdU, the product’s chemistry removes the need for DNA denaturation or harsh cell treatment, adversely affecting sample integrity, cell morphology, image quality, and the ability to multiplex fluorophores. This allows it to be given to animals with minimal toxicity, further, it opens the possibility of live cell/tissue proliferation detection now that DNA denaturation is no longer needed. Click-iT EdU Cell Proliferation Kits can be used for a variety of applications to learn more about these applications, visit our informational pages: fluorescence microscopy, high content screening and flow cytometry.
How does click chemistry help label antibodies?
SiteClick Antibody Labeling Kits use click chemistry for site selective labeling. This is achieved by enzymatic modification of the carbohydrate domain of the antibody to attach an azide to the Fc region of the antibody, preventing fluorophore attachment to the antigen binding sites of the antibody, which renders it ineffective. The enzymatic modification allows for mild reaction conditions that help preserve antibody binding and structure. These features of SiteClick labeling are critical for antibody applications requiring high specificity with reproducible labeling reactions and help ensure that antibody performance and function are preserved after conjugation.
Conclusion
The click chemistry used in our Click-iT EdU and SiteClick products allows for, specific, and gentle labeling of nucleic acids and proteins, which can open exciting possibilities for assaying proliferation in live or fixed samples and help increase the quality and consistency of antibody labelling respectively.
To learn more about all of the Thermo Fisher Scientific cellular imaging offerings, tools and resources, bookmark and explore the latest by visiting the cellular imaging resource center at thermofisher.com/cellularimaging.
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