AI Article Synopsis
- The standard method to measure cells in the S-phase of the cell cycle has involved 5-bromo-2'-deoxyuridine (BrdU) labeling and antibody staining, but it can damage DNA structure.
- An improved method using 5-ethynyl-2'-deoxyuridine (EdU) leverages click chemistry, allowing for more efficient detection of S-phase cells without disrupting the DNA helix.
- This EdU approach preserves DNA structure and cell surface markers, leading to shorter assay times and better reproducibility compared to the BrdU method.
Article Abstract
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.
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| http://dx.doi.org/10.2144/000112812 | DOI Listing |
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