Improved methodology for detecting bromodeoxyuridine in cultured cells and tissue sections by immunocytochemistry.

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  • Author(s): Dover R;Dover R; Patel K
  • Source:
    Histochemistry [Histochemistry] 1994 Nov; Vol. 102 (5), pp. 383-7.
  • Publication Type:
    Journal Article
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: Springer-Verlag Country of Publication: Germany NLM ID: 0411300 Publication Model: Print Cited Medium: Print ISSN: 0301-5564 (Print) Linking ISSN: 03015564 NLM ISO Abbreviation: Histochemistry Subsets: MEDLINE
    • Publication Information:
      Original Publication: Berlin ; New York : Springer-Verlag.
    • Subject Terms:
      Bromodeoxyuridine/*analysis; Animals ; Antibodies, Monoclonal ; Cells, Cultured ; Fibroblasts/chemistry ; Fibroblasts/cytology ; Humans ; Immunohistochemistry ; Male ; Mice ; Microtomy ; Microwaves ; Rats ; Skin/anatomy & histology ; Skin/chemistry ; Skin/cytology ; Staining and Labeling/methods
    • Abstract:
      Detection of DNA synthesizing cells may often be achieved by immunocytochemical detection of bromodeoxyuridine (BrdU), which is rapid and appears to give similar results to those found using tritiated thymidine. However, the methodology for detection of BrdU involves a denaturation or digestion step to allow access of the antibody to BrdU incorporated into single- rather than double-stranded DNA. We wished to determine if microwave treatment could be used to enhance the detection of BrdU without the need for any other digestion/denaturation steps. An important consideration was to investigate whether such treatment produces a similar quantitative result, since BrdU detection is usually assessed on the basis of cell number rather than topographical distribution. We have found that microwave treatment can allow considerably lower antibody concentrations and eliminates the need for any other denaturation step. It also reduces the non-specific background staining found when using monoclonal antibodies on mouse tissue. We have performed cell counts and found that the number of BrdU positive cells remains constant for a range of different immunocytochemical parameters. We also report conditions where immunopositivity is adversely affected by changes in technique and describe the optimised conditions for obtaining reproducible results.
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    • Accession Number:
      0 (Antibodies, Monoclonal)
      G34N38R2N1 (Bromodeoxyuridine)
    • Publication Date:
      Date Created: 19941101 Date Completed: 19950330 Latest Revision: 20190830
    • Publication Date:
      20240829
    • Accession Number:
      10.1007/BF00268909
    • Accession Number:
      7532640