2008/06/01 by Suzanne B. Buck, Jolene A. Bradford, Jolene Bradford +5 · 1 citation
Biochemistry, Genetics and Molecular Biology · Chemistry · Medicine · #Advanced biosensing and bioanalysis techniques #Azide #Biochemistry #Biology #Bromodeoxyuridine #Cell growth #Chemistry #Click Chemistry and Applications #Click chemistry #Combinatorial chemistry #DNA #Deoxyuridine #Molecular biology #Monoclonal and Polyclonal Antibodies Research #Nucleic acid #Nucleoside #Organic chemistry #Thymidine
paper · doi:10.2144/000112812
openalex publication_date 2008/06/01 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/26
The 5-bromo-2'-deoxyuridine (BrdU) labeling of cells followed by antibody staining has been the standard method for direct measurement of cells in the S-phase. Described is an improved method for the detection of S-phase cell cycle progression based upon the application of click chemistry, the copper(I)-catalyzed variant of the Huisgen [3+2] cycloaddition between a terminal alkyne and an azide. 5-ethynyl-2'-deoxyuridine (EdU) is a nucleoside analog of thymidine that is incorporated into DNA during active DNA synthesis, just like BrdU. While the BrdU assay requires harsh chemical or enzymatic disruption of helical DNA structure to allow for direct measurement of cells in the S-phase by the anti-BrdU antibody, the EdU method does not. Elimination of this requirement results in the preservation of helical DNA structure and other cell surface epitopes, decreased assay time, and increased reproducibility.