Assay Principle
Detection of cell proliferation is essential for assessing cell health, determining genotoxicity, or evaluating anticancer drugs.Currently, direct measurement of DNA synthesis is the most accurate method. Nucleoside analogs (e.g., [³H]thymidineor5-bromo-2′-deoxyuridine, BrdU) are incorporated into cells during replication and subsequently detected by autoradiographyorusing anti-BrdU antibodies.
The EdU Cell Proliferation Imaging Analysis Kit (Green Fluorescence) is a novel alternative to the Brd Uassay. EdU(5-ethynyl-2′-deoxyuridine) is a nucleoside analog of thymidine that can be incorporated into newly synthesized DNA in place of thymidine during DNA synthesis. Compared with the BrdU method, the EdU-Click detection is not antibody-based, thus eliminating the need for DNA denaturation (typically performed using hydrochloric acid, heat, or DNase digestion) to detect the incorporated nucleoside. The detection is based on the Click reaction, a copper-catalyzed covalent reaction between anodize and an alkyne, and the entire reaction can be completed within 30 min
Background
The detection of cell proliferation is of utmost importance for assessing cell health, determining genotoxicity or evaluating anticancer drugs. Until now, measuring DNA synthesis directly is most accurate method of doing it, normally performed by incorporation of the nucleoside analog like [3H] thymidine or 5-bromo-2’-deoxyuridine to cells during replication, and then detected or visualized by autoradiography or with an anti-BrdU-antibody respectively.
Proliferating Hela cells detected using Cell Proliferation EdU Image Kit (Green Fluorescence)
