Optimization of the (32)P-Postlabeling/Thin Layer Chromatography Assay ((32)P-TLC) for In Vitro Detection of 8-Oxo-Deoxyguanosine as a Biomarker of Oxidative DNA Damage.
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 (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 over-estimate the level of 8-oxo-dG. In the present study, we optimized the (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 (32)P-postlabeling procedures and were in the range of those determined by chromatography methods (GC-MS, HPLC-MS).
Maatouk
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I
Institut Curie-Section Recherche, Laboratoire Raymond Latarjet, Bâtiment 112, Centre Universitaire, 91405, Orsay Cedex, France.
Bouaïcha
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N
Plessis
M J
MJ
Périn
F
F