Immunological Detection and Quantification of Oxidized Proteins by Labelling with Digoxigenin

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  • BAUTISTA Juan
    Department of Biochemistry, Bromatology and Toxicology, Faculty of Pharmacy, University of Sevilla
  • MATEOS-NEVADO María D.
    Department of Biochemistry, Bromatology and Toxicology, Faculty of Pharmacy, University of Sevilla

書誌事項

タイトル別名
  • Immunological Detection and Quantificat
公開日
1998
資源種別
journal article
DOI
  • 10.1271/bbb.62.419
公開者
公益社団法人 日本農芸化学会

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説明

An immunological assay, based on the digoxigenin/anti-digoxigenin system, was developed to detect and quantify carbonyl moieties that result from oxidative damage to proteins. Bovine serum albumin (BSA) was oxidized by a hydroxyl radical-generating system consisting of ascorbate/Fe(III)/O2. 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 pmole-amount 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.<br>

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