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通过酶联免疫吸附测定(ELISA)或高效液相色谱(HPLC)结合电化学检测法测定氧化性DNA损伤8-氧代鸟嘌呤(8-oxoG)

Measurement of the Oxidative DNA Lesion 8-Oxoguanine (8-oxoG) by ELISA or by High-Performance Liquid Chromatography (HPLC) with Electrochemical Detection.

作者信息

Drake Danielle M, Shapiro Aaron M, Wells Peter G

机构信息

Department of Pharmaceutical Sciences and Centre for Pharmaceutical Oncology, University of Toronto, Toronto, ON, Canada.

Department of Pharmaceutical Sciences, University of Toronto, Toronto, ON, Canada.

出版信息

Methods Mol Biol. 2019;1965:313-328. doi: 10.1007/978-1-4939-9182-2_21.

Abstract

Reactive oxygen species (ROS) can oxidize cellular macromolecules like DNA, causing DNA damage. The most common form of DNA damage is the 8-oxoguanine (8-oxoG) lesion, typically repaired by the base excision repair (BER) pathway, which is initiated by the enzyme oxoguanine glycosylase 1 (OGG1). ROS are produced endogenously and can be enhanced by environmental factors, such as xenobiotics, radiation, and microbial pathogens. As a commonly used biomarker of oxidative damage, 8-oxoG can be measured in two different ways described herein. Commercially available ELISA kits allow for easy detection of the 8-oxoG lesion, while more difficult HPLC assays with UV and electrochemical detection allow for a more definitive identification and quantification of 8-oxoG.

摘要

活性氧(ROS)可氧化细胞大分子如DNA,导致DNA损伤。DNA损伤最常见的形式是8-氧代鸟嘌呤(8-oxoG)损伤,通常由碱基切除修复(BER)途径修复,该途径由氧代鸟嘌呤糖基化酶1(OGG1)启动。ROS是内源性产生的,并且可被环境因素增强,如外源性物质、辐射和微生物病原体。作为氧化损伤的常用生物标志物,8-oxoG可通过本文所述的两种不同方法进行测量。市售的ELISA试剂盒可轻松检测8-oxoG损伤,而更具难度的采用紫外和电化学检测的高效液相色谱法(HPLC)可对8-oxoG进行更明确的鉴定和定量。

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