4-HNE(4-Hydroxynonenal) ELISA Kit (E-EL-0128)

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Universal 4-HNE. During the reaction, Universal 4-HNE in the sample or standard competes with a fixed amount of Universal 4-HNE on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Universal 4-HNE. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Universal 4-HNE in tested samples can be calculated by comparing the OD of the samples to the standard curve.

4-hydroxynonenal (4-HNE) is an alpha,beta-unsaturated hydroxyalkenal with the formula C9H16O2 generated by peroxidation of polyunsaturated fatty acids, particularly arachidonic and linoleic acid, in membrane phospholipids. Because such fatty acids are highly susceptible to attack by reactive oxygen species, it is one of the most abundant products of lipid peroxidation. Its concentration rises markedly during oxidative stress, and its electrophilic structure allows it to form covalent adducts with proteins and DNA. At low concentrations 4-HNE functions as an adaptive signaling molecule that induces antioxidant and detoxification responses, whereas at higher concentrations it is cytotoxic. Modified proteins thus serve both as markers of membrane injury and as mediators of that injury. Measurement of 4-HNE and its protein adducts is widely used as a lipid peroxidation marker in studies of aging, neurodegenerative disease, atherosclerosis, diabetes and toxic liver injury. Quantitation in tissue, plasma and cultured cells assesses membrane damage, screens antioxidant compounds, and supports investigation of how lipid peroxidation products drive inflammation and cell death and of the detoxification of reactive aldehydes.