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.

Abstract:

: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).

journal_name

Toxicol Mech Methods

authors

Maatouk I,Bouaïcha N,Plessis MJ,Périn F

doi

10.1080/15376520600616909

subject

Has Abstract

pub_date

2006-01-01 00:00:00

pages

313-22

issue

6

eissn

1537-6516

issn

1537-6524

journal_volume

16

pub_type

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