Application of Antibodies to 5-Bromodeoxyuridine for the Detection of Cells of Rare Genotype

  • Howard G. Gratzner
Part of the Topics in Chemical Mutagenesis book series (TCM, volume 2)


A current objective of research in the area of environmental mutagenesis is the development of automated methods of assessing in vivo and in vitro genetic damage.(1,2) The primary purpose of automating mutagen detection is to increase the speed and statistical reliability that is afforded by the instrumentation being recruited to this task, flow or image cytometry. In addition, this approach would eliminate the human bias and error characteristic of manual methods. In the case of in vivo assessment of mutagenesis, a method would be available to measure genetic variation in cells that either do not clone or clone at extremely low efficiencies.


Sister Chromatid Unlabeled Cell Rare Genotype Indirect Immunoperoxidase Technique Dansyl Hydrazine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    M. L. Mendelsohn, W. L. Bigbee, E. W. Branscomb, and G. Stamatoyannopaulos, The detection and sorting of rare sickle-hemoglobin containing cells in normal human blood, Flow Cytom. IV, 311–313 (1980).Google Scholar
  2. 2.
    B. Holtkamp, M. Cramer, H. Lemke, and K. Rajewsky, Isolation of a cloned cell line expressing variant H-2K+ by fluorescence-activated cell sorting, Nature 289, 66–68 (1981).PubMedCrossRefGoogle Scholar
  3. 3.
    P. Howard-Flanders, Mutagenesis in mammalian cells, Mutat. Res. 86, 307–327 (1981).PubMedGoogle Scholar
  4. 4.
    Committee 17, Environmental mutagenic hazards, Science 187, 503–514 (1975).CrossRefGoogle Scholar
  5. 5.
    B. N. Ames, Identifying environmental chemicals causing mutations and cancer, Science 204, 587–593 (1979).PubMedCrossRefGoogle Scholar
  6. 6.
    A. Abbondandolo, Prospects for evaluating genetic damage in mammalian cell culture, Mutat. Res. 42, 279–298 (1977).PubMedCrossRefGoogle Scholar
  7. 7.
    P. E. Nute, Th. Papayannopaulou, B. Tatsis, and G. Stamatoyannopoulos, Towards a system for detecting somatic-cell mutations. V. Preparation of fluorescent antibodies to hemoglobin Hasharon, A human a chain variant, J. Immunol. Methods 42, 5–44 (1981).CrossRefGoogle Scholar
  8. 8.
    G. Stamatoyannopoulos, P. E. Nute, Th. Papayannopoulou, T. Mcguire, G. Lim, H. F. Bunn, and D. Ruchnagel, Development of a somatic mutation screening system using Hb mutants. IV. Successful detection of red cells containing the human frameshift mutants Hb Wayne and Hb Cranston using monospecific antibodies, Am. J. Hum. Genet. 32, 484–496 (1980).PubMedGoogle Scholar
  9. 9.
    G. Strauss and J. Albertini, Enumeration of 6-thioguanine-resistant peripheral blood lymphocytes in man as a potential test for somatic cell mutations arising in vivo, Mutation 61, 353–379 (1979).Google Scholar
  10. 10.
    W. Herzenberg and L. Herzenberg, Analysis and separation using the fluorescence activated cell sorter (FACS), in: Handbook of Experimental Immunology (D. M. Wier, ed.), Vol. 2, pp. 221–222, Blackwell, Oxford (1978).Google Scholar
  11. 11.
    S. A. Latt, Microfluorometric determination of deoxyribonucleic acid replication in human metaphase chromosomes, Proc. Natl. Acad. Sci USA 70, 3395–3398 (1973).PubMedCrossRefGoogle Scholar
  12. 12.
    D. E. Swartzendruber, A bromodeoxyuridine (BrdUrd) mithramycin technique for detecting cycling and non-cycling cells by flow microfluorometry, Exp. Cell Res. 109, 439–443 (1977).PubMedCrossRefGoogle Scholar
  13. 13.
    E. E. Furth, W. G. Thilly, B. W. Penman, H. L. Liber, and W. M. Rand, Quantitative assay for mutation in diploid human fibroblasts using microtiter plates, Anal. Biochem. 110, 1–8 (1981).PubMedCrossRefGoogle Scholar
  14. 14.
    G. Kohler and C. Milstein, Continuous cultures of fused cells secreting antibody of predefined specificity, Nature 256, 495–497 (1975).PubMedCrossRefGoogle Scholar
  15. 15.
    H. G. Gratzner, Monoclonal antibody to 5-Bromo and 5-Iododeoxyuridine: a new reagent for detection of DNA replication, Science, 28, 474–475 (1982).CrossRefGoogle Scholar
  16. 16.
    H. G. Gratzner, A. Pollack, D. J. Ingram, and R. C. Leif, Deoxyribonucleic acid replication in single cells and chromosomes by immunologic techniques, J. Histochem. Cytochem. 24, 34–39 (1976).PubMedCrossRefGoogle Scholar
  17. 17.
    B. F. Erlanger and S. M. Beiser, Antibodies specific for ribonucleosides and ribonucleotides and their reaction with DNA, Proc. Natl. Acad. Sci USA 52, 68–74 (1964).PubMedCrossRefGoogle Scholar
  18. 18.
    H. G. Gratzner, R. C. Leif, D. J. Ingram, and A. Castro, The use of antibodies specific for bromodeoxyuridine for the immunofluorescent determination of DNA replication in cells and chromosomes, Exp. Cell Res. 95, 88–93 (1975).PubMedCrossRefGoogle Scholar
  19. 19.
    O. A. Sternberger, Immunocytochemistry, Prentice-Hall, Englewood Cliffs, New Jersey (1974).Google Scholar
  20. 20.
    M. Shulman, C. D. Wilde, and G. Kohler, A better cell line for making hybridomas secreting specific antibodies, Nature 276, 269–270 (1978).PubMedCrossRefGoogle Scholar
  21. 21.
    M. L. Gefter, D. L. Margulies, and M. Scharff, A simple method for the polyethylene glycolpromoted hybridization of mouse myeloma cells, Somatic Cell Genet. 3, 231–236 (1977).PubMedCrossRefGoogle Scholar
  22. 22.
    Anon, Preparation of antigen coated plates for use in enzyme-linked immunoassays. Hybrlines I(5), 5 (1980).Google Scholar
  23. 23.
    H. G. Gratzner, N. Ettinger, and D. J. Ingram, Immunochemical studies of 5-bromodeoxyuridine, Res. Commun. Chem. Pathol. Pharmacol. 20, 539–598 (1978).PubMedGoogle Scholar
  24. 23a.
    F. Dolbeare, H. G. Gratzner, M. Pallavicini, and J. W. Gray, Flow cytometric measurement of total DM content and incorporated bromodeoxyuridine. Proc. Nat. Acad. Sci USH (1984), in press.Google Scholar
  25. 24.
    P. Laurila, I. Virtanen, J. Wartiovaara, and S. Stenman, Fluorescent antibodies and lectins stain intracellular structures in fixed cells treated with non-ionic detergent, J. Histochem. Cytochem. 26, 251–257 (1978).PubMedCrossRefGoogle Scholar
  26. 25.
    H. G. Gratzner and R. C. Leif, An immunofluorescent method for monitoring DNA synthesis by flow cytometry. Cytometry 1, 385–389 (1981).PubMedCrossRefGoogle Scholar
  27. 26.
    R. B. Painter, Rapid test to detect agents that damage DNA, Nature 265, 650–651 (1977).PubMedCrossRefGoogle Scholar
  28. 27.
    C. Clark, The mutagenic specificities of pentachloronitrobenzene and captan, two environmental mutagens, Mutat. Res. 11, 247–248 (1971).Google Scholar
  29. 28.
    L. L. Wotring and J. L. Roti Roti, Thioguanine-induced S and G2 blocks and their significance to the mechanism of toxicity, Cancer Res. 40, 1458–1562 (1980).PubMedGoogle Scholar
  30. 29.
    R. H. Kennett, T. J. Mckearn, and K. B. Bechtol, eds., Monoclonal Antibodies. Hybridomas: A New Dimension in Biological Analyses, Plenum Press, New York (1980).Google Scholar

Copyright information

© Plenum Press, New York 1984

Authors and Affiliations

  • Howard G. Gratzner
    • 1
  1. 1.Institute for Cell Analysis and Department of MedicineUniversity of Miami School of MedicineMiamiUSA

Personalised recommendations