Automated determination of 8-OHdG in cells and tissue via immunofluorescence using a specially created antibody

使用专门创建的抗体通过免疫荧光自动测定细胞和组织中的 8-OHdG

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作者:Tobias Jung, Nicole Findik, Bianca Hartmann, Katja Hanack, Kai Grossmann, Dirk Roggenbuck, Marc Wegmann, René Mantke, Markus Deckert, Tilman Grune

Abstract

Despite powerful DNA repair systems, oxidative damage/modification to DNA is an inevitable side effect of metabolism, ionizing radiation, lifestyle habits, inflammatory pathologies such as type-2 diabetes or metabolic syndrome, cancer and natural aging. One of the most common oxidative DNA modifications is 8-OHdG (8‑hydroxy-2'-deoxyguanosine), which is the most widely used marker in research and clinical diagnostics. 8-OHdG is easily and specifically detectable in various samples such as urine, plasma, cells and tissues via a large variety of methods like ELISA, HPLC, chromatographic methods, and immunochemistry. Formed by oxidation of guanine and being representative for the degree of DNA damage, 8-OHdG can be also used as biomarker for risk assessment of various cancers as well as degenerative diseases. Here, we present a highly specific, self-developed 8-OHdG antibody in successful comparison to a commercially one, tested in cells (FF95, HCT116, and HT22) and intestinal tissue, focusing on automatized evaluation via fluorescence/confocal microscopy.

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