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R&D Systems now offers a Microplate-based TBARS Parameter Assay Kit (Catalog # KGE013) that does not require a boiling step. This kit is designed to measure Thiobarbituric Acid Reactive Substances (TBARS) generated during oxidative stress. The oxidative degradation of lipids by reactive oxygen species (ROS), called lipid peroxidation, results in the formation of highly reactive and unstable lipid peroxides. Decomposition of lipid peroxides results in the formation of TBARS, including malondialdehyde (MDA). Measuring TBARS levels offers a convenient method of determining the relative lipid peroxide content in a sample. The TBARS Parameter Assay Kit quantifies TBARS levels in cell culture supernates, cell lysates, serum, plasma, and urine by setting up a reaction between MDA and two molecules of 2-thiobarbituric acid (TBA). In the presence of heat and acid, MDA reacts with TBA to produce a colored end product that can be measured. Intensity of the color corresponds to the level of lipid peroxidation in the sample.
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| Assay Principle for the TBARS Parameter Kit. Acid-treated samples and standards, followed by the TBA reagent, are added to the included 96-well microplate (Step 1). The microplate is then incubated at 45-50 °C for 2-3 hours, during which time the MDA in the sample reacts with the TBA reagent to produce a colored end product (Step 2). The microplate is read at 532 nm and the intensity of the color corresponds to the level of lipid peroxidation in the sample (Step 3). |
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Measurement of TBARS/MDA in Biological Fluids. Samples were gathered from apparently healthy volunteers. Human serum (N=38), plasma with EDTA (N=38) or heparin (N=38), and urine (N=20) were assayed for TBARS levels using the TBARS Parameter Assay Kit (Catalog # KGE013). |