Description
• Quick 30 minute assessment for oxidative stress
• User friendly protocol uses smaller reaction volumes in a 96-well format
• Does not require glass tubes or marbles
The TBARS (Thiobarbituric Acid Reactive Substances) assay is well-established for screening and monitoring lipid peroxidation. MDA forms a 1:2 adduct with thiobarbituric acid; the MDA-TBA adduct can then be measured. Our OxiSelect TBARS Assay Kit provides a much more user-friendly protocol to measure the MDA-TBA adduct. Reaction volumes are much smaller than the traditional assay, so much less sample is required. Also, reactions can be performed in standard polypropylene tubes - no glass tubes or glass marbles are required. Important Note: MDA adducts are not stable long term. For best results test all samples immediately upon collection, or freeze them at -80 degree C for up to one month. MDA may be degraded in samples that have been frozen for longer periods; in such cases more reliable results may be obtained from more stable markers of oxidative stress such as protein carbonyl, 8-OHdG or 4-HNE.
Introduction: Lipid peroxidation is a well-defined mechanism of cellular damage in animals and plants. Lipid peroxides are unstable indicators of oxidative stress in cells that decompose to form more complex and reactive compounds such as Malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE), natural bi-products of lipid peroxidation. Oxidative modification of lipids can be induced in vitro by a wide array of pro-oxidant agents and occurs in vivo during aging and in certain disease conditions. Measuring the end products of lipid peroxidation is one of the most widely accepted assays for oxidative damage. These aldehydic secondary products of lipid peroxidation are generally accepted markers of oxidative stress.Thiobarbituric Acid Reactive Substances (TBARS) is a well-established assay for screening and monitoring lipid peroxidation. The rapid and easy protocol has been modified by researchers in the evaluation of drugs, food, as well as human and animal tissue samples. MDA forms a 1:2 adduct with thiobarbituric acid (Figure 1). The MDA-TBA adduct formed from the reaction of MDA in samples with TBA can be measured colorimetrically or fluorometrically. TBARS levels are determined from a Malondialdehyde equivalence standard.The TBARS Assay has provided relevant information concerning free radical activity in disease states and measurement of many compounds anti-oxidant characteristics. Although the specificity of TBARS toward compounds other than MDA has been controversial, the assay continues to be the most widely employed format for monitoring lipid peroxidation. Lipids with higher degrees of unsaturated bonds produce higher TBARS values. Interfering soluble TBARS can be minimized if lipoprotein fractions are first acid precipitated from samples. Biological samples may contain a mixture of thiobarbituric acid reactive substances such as hydroperoxides and aldehydes, which increase in response to oxidative stress. If excessively high TBARS values are obtained, a more specific assay such as HPLC should be employed.The OxiSelect TBARS Assay Kit offers a simple, reproducible, and consistent system for the detection of lipid peroxidation in urine, plasma, serum, lysates, and tissue homogenates. This kit includes an MDA standard for use as a positive control. Each kit provides sufficient reagents to perform 200 tests including standard curve and unknown samples