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  • 5-Ethynyl-2'-deoxycytidine as a new agent for DNA labeling: detection of proliferating cells.

5-Ethynyl-2'-deoxycytidine as a new agent for DNA labeling: detection of proliferating cells.

Analytical biochemistry (2011-06-21)
Dezhong Qu, Guoxing Wang, Zhe Wang, Li Zhou, Weilin Chi, Shujie Cong, Xiaoshuai Ren, Peizhou Liang, Biliang Zhang
ABSTRACT

The labeling of newly synthesized DNA in cells to identify cell proliferation is an important experimental technique. The most accurate methods incorporate [(3)H]thymidine or 5-bromo-2'-deoxyruidine (BrdU) into dividing cells during S phase, which is subsequently detected by autoradiography or immunohistochemistry, directly measuring the newly synthesized DNA. Recently, a novel method was developed to detect DNA synthesis in proliferating cells based on a novel thymidine analog, 5-ethynyl-2'-deoxyuridine (EdU). EdU is incorporated into DNA and subsequently detected with a fluorescent azide via "click" chemistry. This novel technique is highly sensitive and does not require DNA denaturation. However, it was also found that EdU exhibits time-dependent inhibition effects on cell growth. Therefore, here we report a novel deoxycytidine analog, 5-ethynyl-2'-deoxycytidine (EdC), that can be used to detect DNA synthesis in vitro and in vivo at a similar sensitivity level compared with EdU. Furthermore, the EdC-induced cytotoxicity is much less than that of EdU when combined with thymidine. This will be a potential application for the long-term detection of proliferating cells.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
5-Ethynyl-2′-deoxycytidine, (EdC), AldrichCPR
Sigma-Aldrich
5-Ethynyl-2′-deoxyuridine, 95%