Cell proliferation assays are widely used in the evaluation of cell viability, genotoxicity, and the effect of antitumor drugs. Direct detection of DNA synthesis in cells is considered to be the most accurate method for detecting cell proliferation. The initial widely used method for detecting DNA synthesis in cells was the radiolabeled nucleoside incorporation method, but this method was greatly limited due to radioactive contamination and the difficulty of single-cell detection, and was gradually replaced by the BrdU method based on antibody detection. The BrdU method has many steps and requires the use of BrdU antibody, which has many influencing factors and poor stability.
EdU method is based on EdU incorporation and subsequent click reaction, without the use of antibodies, convenient operation and high detection sensitivity. It is a new method upgraded on the basis of BrdU method and will gradually replace BrdU method. EdU (5-ethynyl-2-deoxyuridine), is a thymidine analog, EdU can replace thymidine in the process of DNA synthesis to incorporate into new in synthetic DNA. On the other hand, the acetylene group on EdU can react with fluorescently labeled small molecule azide probes (such as FITC Azide, Elab Fluor® 488 Azide, Elab Fluor® 594 Azide, Elab Fluor® 647 Azide) through the catalysis of monovalent copper ions to form a stable triazole ring. This reaction is very rapid and is called the click reaction. Through the click reaction, the newly synthesized DNA is labeled with the corresponding fluorescent probe, so that the proliferating cells can be detected using the appropriate fluorescent detection equipment.
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