Lipid peroxidation is a well-established mechanism of cellular injury in both plants and animals, and is used as an indicator of oxidative stress in cells and tissues. Lipid peroxides are unstable and decompose to form a complex series of compounds including reactive carbonyl compounds. Polyunsaturated fatty acid peroxides generate malondialdehyde (MDA) and 4-hydroxyalkenals (HAE) upon decomposition.
Measurement of malondialdehyde and 4-hydroxyalkenals has been used as an indicator of lipid peroxidation. This method is designed to assay either MDA alone (in hydrochloric acid) or MDA in combination with 4-hydroxyalkenals (in methanesulfonic acid.)
Principle of the Assay:
This assay is based on the reaction of a chromogenic reagent, N-methyl-2-phenylindole (R1), with MDA and 4-hydroxyalkenals at 45 degrees C. One molecule of either MDA or 4-hydroxyalkenal reacts with 2 molecules of reagent R1 to yield a stable chromophore with maximal absorbance at 586nm.
L2496-30A: Reagent R1, 3 x18ml. Supplied as a liquid , N-methyl-2-phenylindole in acetonitrile
L2496-30B: Reagent R2, 1x16.5ml. Supplied as a liquid, Methanesulfonic acid (MSA)
L2496-30C: MDA Standard (S2), 1x1ml. Supplied as a liquid, 1,1,3,3-Tetramethoxypropane in Tris-HCl
L2496-30D: Diluent, 1x30ml. Supplied as a liquid Ferric Iron in Methanol
Storage and Stability:
Store all components at 4 degrees C. Stable for at least 6 months