Here, using the embryonic mouse cerebral cortex as a model system, we describe a simplified dual thymidine analogue protocol using BrdU and EdU that can be used to measure T s and T c. The advantage of this protocol over cumulative labeling approaches is that only a single time-point is required for measurement.

Dual pulse labeling of cell proliferation traditionally uses BrdU immunocytochemistry with iododeoxyuridine (IdU) or chlorodeoxyuridine (CldU), using multiple BrdU antibodies of different clones that cross-react with IdU and CldU for detection [7].

With the availability of new highly specific anti-BrdU antibodies, the BrdU labeling technique can be combined with click chemistry detection for a simplified method of dual pulse labeling.

Thymidine analogues can be detected by a variety of methods including radio-detection for tritiated thymidine, immuno-detection for bromo-deoxyuridine (BrdU), chloro-deoxyuridine (CldU) and iodo-deoxyuridine (IdU), and chemical detection for ethinyl-deoxyuridine (EdU).

Incorporation of the thymidine analog 5′-bromo-2′-deoxyuridine (BrdU) has been established as a popular assay for determining cell proliferation rates in a wide variety of species, ranging from plants to mammalian cells (Cecchini et al. 2012, Nagar et al. 2002). One common assay set-up is to supplement the culture media of growing cells with BrdU.

BrdU • 5-bromo-2’-deoxyuridine (BrdU) is a thymidine analog that is incorporated into newly synthesized DNA • Labels proliferating and daughter cells • Detected by staining with anti-BrdU antibodies • Can be used to accurately quantify the percentage of cells in G1, S, and G2/M, and trace cell cycle kinetics

Feb 14, 2018 · An additional benefit of this protocol over existing dual-analog approaches (CldU/IdU) is the antibody-free detection of EdU and the acid-free detection of BrdU, processes allowing for the parallel use of specific antibodies so as to measure the cell-cycle in immunologically defined cellular subpopulations.

Jan 16, 2024 · Overall, our study finds that BrdU/IdU can activate the DNA damage repair pathway (HRR), leading to increased histone acetylation and genome-wide DNA demethylation, regulating somatic cell reprogramming. This offers valuable insights into mechanisms underlying cell fate transition.

Feb 14, 2018 · 5-Bromo-2'-deoxyuridine (BrdU) and 2'-deoxy-5-ethynyluridine (EdU) are widely used as markers of replicated DNA. While BrdU is detected using antibodies, the click reaction typically with ...

Apr 22, 2019 · Using this method, we detect 5-bromodeoxyuridine (BrdU) incorporated by Saccharomyces cerevisiae to reveal, at a genomic scale and on single molecules, the DNA sequences replicated during a...