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BrdU Incorporation Assay to Analyze the Entry into S Phase

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Cell-Cycle Synchronization

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2579))

Abstract

5-Bromo-2′-deoxyuridine (bromodeoxyuridine (BrdU)), a modified nucleotide and analog of thymidine, is commonly used for detecting proliferating cells. For detection, an anti-BrdU antibody (probe) with a fluorescent dye is applied to bind the BrdU label after DNA denaturation. In this protocol, we provide the BrdU labeling method for both in vitro and in vivo studies, along with immunocytochemistry (ICC)/immunofluorescence (IF) and immunohistochemistry (IHC) staining procedures, respectively. Multicolor staining is also presented as an option to detect the co-distribution of two or multiple antigens in the same sample, making it possible to visualize the location of different molecules at the same time.

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Acknowledgments

This work was supported by Atomic Energy of Canada Limited’s Federal Nuclear Science & Technology Program and CANDU Owners Groups (COG) Strategic Research and Development (SRD) program.

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Correspondence to Yi Wang .

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Yu, J., Wang, Z., Wang, Y. (2022). BrdU Incorporation Assay to Analyze the Entry into S Phase. In: Wang, Z. (eds) Cell-Cycle Synchronization. Methods in Molecular Biology, vol 2579. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2736-5_16

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  • DOI: https://doi.org/10.1007/978-1-0716-2736-5_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2735-8

  • Online ISBN: 978-1-0716-2736-5

  • eBook Packages: Springer Protocols

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