Millipore Technical Publications |
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Poster: Methods For The Detection of Oxidative Stress | ||
| Lit No: | ps1040en00 | |
| Year: | 2008 | |
Introduction
Oxidative stress is a cellular disorder caused by an excess of free radicals which can occur because of increased oxidant levels, decreased anti-oxidant amounts, or failure to repair oxidative damage induced by reactive oxygen species (ROS). This condition has been linked to the pathogenesis of a number of disease states including atherosclerosis, ischemia, cancer, rheumatoid arthritis, and neurodegeneration. ROS are free radicals, reactive molecules containing oxygen, and include superoxides, hydroxyl radicals, hydrogen peroxides, nitric oxides, peroxynitrites, and singlet oxygens. ROS molecules are capable of chemically modifying proteins, lipids, and nucleic acids which potentially can alter or impair their natural biological function. Multiple approaches for detection of modifications are needed due to the variety of ROS molecules that exist as well as the variety of alterations they can impart. The most common approach is to detect a modification or protein involved in processing using an antibody. This is typically done by western blot analysis or IHC. Approaches to detect modifications to DNA (anti-8 hydroxydeoxyguanosine or anti-8-oxoguanine), lipids (anti-neuroketal, anti-4- hydroxynonenal), or proteins (anti-SOD1, anti-ntirotyrosine, anti-nNOS, anti- HIF1a, and more) are commonly utilized. Another method frequently employed to detect ROS modifications is to chemically attach a molecule to a particular group and subsequently detect this extension. Utilizing this methodology we illustrate detection and quantification of carbonyl modifications using the OxyBLOTTM and OxyELISATM assays. Results for each of these methodologies are given as well as discussions about advantages and disadvantages of each. This will allow researchers to decide how best to detect ROS-induced modifications based on their particular needs ranging in scope from molecular to single cell to system wide.Click the PDF above for the full poster.