Immunological Detection and Quantification of Oxidized Proteins by Labelling with Digoxigenin

概要

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An immunological assay, based on the digoxigenin/antidigoxigenin system, was developed to detect and quantify carbonyl moieties that result from oxidatie damage to proteins.Bovine serum albumin (BSA)was oxidized by a hdroxyl radical-generating system consisting of ascorbate/Fe(III)/O_2.The resulting albumin-derived carbonyls were labelled with digoxigenin-hydrazide and detected by dot blotting with an anti-digoxigenin antibody conjugated to alkaline phosphatase.Quantification was carried out by a densitometric analysi.This system allows the detection of a pmoleamount of carbonyl groups on blots.The assay covers a range of sensitivity from 1.26 to 126 pmoles.Another feature of this method is its application to a complex protein mixture(homogenate)to analyze the oxidative status of individual proteins, as are shown for intestinal brush border membrane homogenate of a rat.
社団法人日本農芸化学会の論文
1998-03-23