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Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric) (ab118970) provides a convenient tool for sensitive detection of the malondialdehyde (MDA) present in a variety of samples. MDA, together with 4-hydroxynonenal (4-HNE), is a natural bi-product of lipid peroxidation and its quantification is generally used as marker for lipid peroxidation. The MDA in the sample reacts with thiobarbituric acid (TBA) to generate a MDA-TBA adduct. The MDA-TBA adduct can be easily quantified colorimetrically (OD = 532 nm) or fluorometrically (Ex/Em = 532/553 nm). This assay detects MDA levels as low as 1 nmol/well colorimetrically and 0.1 nmol/well fluorometrically.
The MDA assay is also refered to as a TBARS assay.
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Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.
Lipid peroxidation refers to the oxidative degradation of lipids. In this process free radicals take electrons from the lipids (generally in cell membranes), resulting in cell damage. Quantification of lipid peroxidation is essential to assess oxidative stress in pathophysiological processes. Lipid peroxidation forms reactive aldehydes such as malondialdehyde (MDA) and 4-hydroxynonenal (4- HNE) as natural bi-products. Measuring the end products of lipid peroxidation is one of the most widely accepted assays for oxidative damage.
|BHT (100X)||Purple||1 x 1ml|
|MDA Lysis Buffer||WM||1 x 25ml|
|MDA Standard (4.17M)||Yellow||1 x 100µl|
|Phosphotungstic Acid Solution||NM||1 x 12.5ml|
10 mg of tissue were homogenized on ice in 300 μL of MDA lysis buffer, then centrifuged (13,000 × g, 10 min) to remove insoluble materials. 10 μL of plasma were mixed with 500 μL of 42 mM H2SO4 and 125 μL of phosphotungstic acid solution at room temperature for 5 min. After centrifuging (13,000 × g, 3 min), the pellet was re-suspended on ice with 100 μL of double-distilled H2O. Then, 200 μL of solution and 600 μL of 2-thiobarbituric acid solution were incubated at 95°C for 60 min, before cooling to room temperature in the ice bath for 10 min. The intensity of absorbance at 532 nm was proportional to the MDA level.
Typical MDA standard calibration curve using colorimetric reading.
Typical MDA standard calibration curve using fluorometric reading.
Measurement of MDA in human plasma (20 µl) and rat liver lysate (10 mg).
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