An improved immunocytochemical procedure for high‐sensitivity detection of incorporated bromodeoxyuridine

Wolfgang Beisker, Frank Dolbeare, Joe W. Gray

Research output: Contribution to journalArticlepeer-review

126 Scopus citations


This report describes an improved immunochemical procedure to stain cells in suspension for incorporated bromodeoxyuridine (BrdUrd) and total DNA content. The procedure consists of five steps: (1) chromatin proteins are extracted by treating with 0.1 M HCl and 0.7% Triton X‐100 to facilitate DNA denaturation and to minimize nonspecific staining; (2) cellular DNA is denatured by heating to 100°C in distilled water; (3) BrdUrd in single‐stranded DNA (ssDNA) is stained using an immunochemical procedure; (4) autofluorescence is reduced using sodium borohydride (NaBH4); and (5) DNA is stained with the fluorescent dye propidium iodide. With this procedure, the BrdUrd incorporated by CHO cells during periods as short as a few seconds can be detected using flow cytometry. In addition, the stoichiometry of the immunofluorescent staining procedure is high.

Original languageEnglish (US)
Pages (from-to)235-239
Number of pages5
Issue number2
StatePublished - Mar 1987
Externally publishedYes


  • Anti‐bromodeoxyuridine
  • DNA
  • flow cytometry
  • stoichiometry
  • thermal denaturation

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology


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