The use of antibody specific for bromodeoxyuridine for the immunofluorescent determination of DNA replication in single cells and chromosomes

https://doi.org/10.1016/0014-4827(75)90612-6Get rights and content

Abstract

Antibodies produced against BSA-bromouridine react with bromodeoxyuridine (BUdR) alone, or when incorporated into single-stranded DNA. This was shown by double-diffusion and microcomplement fixation techniques. The antibody, purified by affinity chromatography, can be used for immunofluorescent analysis of DNA replication in single cells, as an alternative to autoradiography with [3H]thymidine. Pulses of BUdR, of 30 min duration, are sufficient for the immunofluorescent detection of DNA synthesis. In addition, the antibody can be used to detect sister chromatid exchanges by the use of the immunoperoxidase technique.

References (17)

  • JG Gall et al.
  • TT Puck et al.

    Biophys j

    (1963)
  • H Quastler et al.

    Exp cell res

    (1959)
  • DM Holm et al.

    Exp cell res

    (1973)
  • D Eichler et al.

    Biochim biophys acta

    (1974)
  • VG Dev et al.

    Exp cell res

    (1972)
  • BW Lubit et al.

    Exp cell res

    (1974)
  • H Kato

    Exp cell res

    (1974)
There are more references available in the full text version of this article.

Cited by (127)

  • Recent advances in nucleotide analogue-based techniques for tracking dividing stem cells: An overview

    2021, Journal of Biological Chemistry
    Citation Excerpt :

    The major disadvantages of 3H-thymidine are handling of a radioactive substance and the use of the time-consuming autoradiography method for detection. Detection of 5-bromo-2′-deoxyuridine (BrdU) (Table 1), a synthetic nucleoside analogue of thymidine, is an alternative technique for the determination of DNA replication and has overcome these disadvantages (12, 13). BrdU incorporated into DNA is recognized by a specific polyclonal or monoclonal antibody produced against bromouridine or iododeoxyuridine complexed to a carrier protein such as bovine serum albumin.

  • Peptidoglycan accelerates granulopoiesis through a TLR2- and MyD88-dependent pathway

    2017, Biochemical and Biophysical Research Communications
  • Isolating dividing neural and brain tumour cells for gene expression profiling

    2016, Journal of Neuroscience Methods
    Citation Excerpt :

    Thymidine analogues are the method of choice for labelling dividing cells in anatomical studies (Cavanagh et al., 2011; Chehrehasa et al., 2009; Gratzner et al., 1975; Salic and Mitchison, 2008).

View all citing articles on Scopus

This work was supported by NCI Contract, NO1-CB-43962 and NO1-CB-33861, and by General Research Support Grant, NIH-5501-RR-05690-03, NIH Grant, CA13441 and by grant 884M from the Council of Tobacco Research-USA Inc.

View full text