Immuno-slot blot assay for detection of UVR-mediated DNA damage

Methods Mol Biol. 2012:920:163-75. doi: 10.1007/978-1-61779-998-3_12.

Abstract

Solar ultraviolet radiation (UVR), through the formation of DNA photolesions, is the primary cause of most skin cancers. A better understanding of the mechanisms of UVR-induced DNA damage may help prevent skin cancer and this may be achieved using methods to quantify DNA damage. The immuno-slot blot (ISB) method is routinely used for detection and quantification of any heat- and alkali-stable DNA adducts for which a sufficiently specific monoclonal antibody is available. The main steps in ISB are fragmentation and denaturation of the DNA, immobilization of DNA to a nitrocellulose filter, incubation with primary antibody against a specific DNA adduct, incubation with an enzyme-linked secondary antibody and finally chemiluminescence detection and quantification of the DNA adducts.

MeSH terms

  • Animals
  • Calibration
  • Cattle
  • DNA / genetics
  • DNA / isolation & purification
  • DNA / metabolism
  • DNA Damage*
  • Deoxyguanosine / metabolism
  • Immunoblotting / methods*
  • Malondialdehyde / metabolism
  • Ultraviolet Rays / adverse effects*

Substances

  • Malondialdehyde
  • DNA
  • Deoxyguanosine