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.
Both MDA and HNE have been shown to be capable of binding to proteins and forming stable adducts, also termed advanced lipid peroxidation end products. These modifications of proteins by MDA or HNE can cause both structural and functional changes of oxidized proteins.
Cell Biolabs’ HNE-His Adduct ELISA Kit is an enzyme immunoassay developed for rapid detection and quantitation of HNE-His protein adducts. The quantity of HNE-His adduct in protein samples is determined by comparing its absorbance with that of a known HNE-BSA standard curve. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown protein samples.
Assay Principle
BSA standards or protein samples (10 μg/mL) are adsorbed onto a 96-well plate for 2 hrs at 37ºC. The HNE-protein adducts present in the sample or standard are probed with an anti-HNE-His antibody, followed by an HRP conjugated secondary antibody. The HNE-protein adducts content in an unknown sample is determined by comparing with a standard curve that is prepared from predetermined HNE-BSA standards.
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