Optimization of the super(32)P-Postlabeling/Thin Layer Chromatography Assay ( super(32)P-TLC) for In Vitro Detection of 8-Oxo-Deoxyguanosine as a Biomarker of Oxidative DNA Damage

• Published in: Toxicology mechanisms and methods Vol. 16; no. 6; pp. 313 - 322
• Main Authors: Maatouk, I, Bouaicha, N, Plessis, MJ, Perin, F
• Format: Journal Article
• Language: English
• Published: 01.08.2006
• ISSN: 1537-6516
• DOI: 10.1080/15376520600616909

During the last years, much attention was focused on the measurement of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG) as a marker of oxidative DNA damage. Among various analytical techniques, the super(32)P-postlabeling assay has been applied in determination of 8-oxo-dG. However, artefactual DNA oxidation could take place during the work-up procedures leading to overestimate the level of 8-oxo-dG. In the present study, we optimized the super(32)P-postlabelling assay with thin layer chromatography to measure 8-oxo-dG in standard samples of 8-oxo-dG, calf thymus DNA and primary cultured rat hepatocytes. The background levels of 8-oxo-dG in calf thymus DNA and in primary cultured rat hepatocytes were lesser than those determined by the previously described super(32)P-postlabeling procedures and were in the range of those determined by chromatography methods (GC-MS, HPLC-MS).