Quantitative immunohistochemical determination of 8-hydroxy-2'-deoxyguanosine by a monoclonal antibody N45.1: its application to ferric nitrilotriacetate-induced renal carcinogenesis model

Lab Invest. 1997 Mar;76(3):365-74.


The DNA base-modified product 8-hydroxy-2'-deoxyguanosine (8-OHdG) is one of the most commonly used markers for the evaluation of oxidative DNA damage. A monoclonal antibody specific for 8-OHdG (N45.1) was characterized and applied in quantitative immunohistochemistry. N45.1 recognized both the modified base and deoxyribose structure of 8-OHdG and required a concentration two orders higher of 8-hydroxyguanosine as a competitor in the ELISA. In addition, N45.1 did not cross-react with the original four deoxyribonucleosides, other DNA base-modified products such as 8-hydroxy-2'-deoxyadenosine and O6-methyl-2'-deoxyguanosine, or urine components such as uric acid, creatine, and creatinine. A ferric nitrilotriacetate-induced rat renal carcinogenesis model was used for the evaluation of quantitative immunohistochemistry. The 8-OHdG index of quantitative immunohistochemistry, as analyzed by NIH image freeware, correlated reasonably well with the 8-OHdG amount determined by high-performance liquid chromatography with an electrochemical detector-except for a difference in peak time, which could be attributed to the selection of target location. The present method has advantages over the high-performance liquid chromatography/electrochemical detector, gas chromatography/mass spectrometry, and 32P-postlabeling methods in that it allows localization of 8-OHdG to be specified without the risk of artifactual production of 8-OHdG during the DNA extraction and hydrolytic processes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 8-Hydroxy-2'-Deoxyguanosine
  • Animals
  • Antibodies, Monoclonal
  • Antibody Specificity
  • Carcinogens*
  • Chromatography, High Pressure Liquid
  • DNA Damage*
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / analysis
  • Ferric Compounds*
  • Immunohistochemistry / methods
  • Kidney / drug effects
  • Kidney / pathology*
  • Kidney Neoplasms / chemically induced
  • Kidney Neoplasms / pathology*
  • Male
  • Models, Biological
  • Nitrilotriacetic Acid / analogs & derivatives*
  • Oxidative Stress
  • Rats
  • Rats, Wistar


  • Antibodies, Monoclonal
  • Carcinogens
  • Ferric Compounds
  • 8-Hydroxy-2'-Deoxyguanosine
  • Deoxyguanosine
  • Nitrilotriacetic Acid
  • ferric nitrilotriacetate