Immunochemical detection of DNA in alkaline sucrose gradient fractions

J. Vijg, E. Mullaart, L. Roza, R. A. Baan, P. H.M. Lohman

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

A procedure for the detection of non-radioactive DNA in alkaline sucrose gradients is described. This method consists of the following steps: (1) fractionation of the DNA-containing gradients into microtiter plates, neutralization and overnight adsorption; (2) covalent labelling of the guanine bases of the adsorbed DNA by reaction with N-acetoxy-2-acetylaminofluorene; and (3) determination of the gradient profile by means of an enzyme-linked immunosorbent assay using antibodies with high affinity for dG-AAF. The method has been found suitable for the rapid and sensitive determination of gamma-ray induced single-strand breaks and UV-induced pyrimidine dimers in mammalian cells in vitro. In addition, it has been shown that the method can be used for the determination of pyrimidine dimers induced in skin cells in vivo.

Original languageEnglish (US)
Pages (from-to)53-58
Number of pages6
JournalJournal of Immunological Methods
Volume91
Issue number1
DOIs
StatePublished - Jul 11 1986
Externally publishedYes

Keywords

  • Alkaline sucrose gradient
  • Automated assay
  • DNA
  • DNA repair
  • Immunochemical method
  • Pyrimidine dimer

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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