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Measurement of 4-Hydroxynonenal (4-HNE) Protein Adducts by ELISA

Part of the Methods in Molecular Biology book series (MIMB,volume 1990)


Enzyme linked immunosorbent assay (ELISA) is a widely used technique for the measurement of antigens and antibodies alike. We describe here procedures, indirect ELISA and sandwich ELISA for the detection of 4-hydroxynonenal protein adducts. These adducts are stable compounds formed within cells and bodily fluids under conditions of oxidative stress. They can act as sensitive biomarkers of oxidative stress and are directly linked to disease pathology.

Key words

  • 4-Hydroxynonenal
  • 4-HNE
  • Anti-HNE antibodies
  • HRP-labeled antibodies
  • Dot blot
  • Indirect ELISA
  • Sandwich ELISA

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  • DOI: 10.1007/978-1-4939-9463-2_4
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  1. Petersen DR, Doorn JA (2004) Reactions of 4-hydroxynonenal with proteins and cellular targets. Free Radic Biol Med 37:937–945

    CAS  CrossRef  Google Scholar 

  2. Lovell MA, Ehmann WD, Mattson MP, Markesbery WR (1997) Elevated 4-hydroxynonenal in ventricular fluid in Alzheimer’s disease. Neurobiol Aging 18:457–461

    CAS  CrossRef  Google Scholar 

  3. Kamimura S, Gaal K, Britton RS, Bacon BR, Triadafilopoulos G, Tsukamoto H (1992) Increased 4-hydroxynonenal levels in experimental alcoholic liver disease: association of lipid peroxidation with liver fibrogenesis. Hepatology 16:448–453

    CAS  CrossRef  Google Scholar 

  4. Shoeb M, Ansari NH, Srivastava SK, Ramana KV (2014) 4-hydroxynonenal in the pathogenesis and progression of human diseases. Curr Med Chem 21:230–237

    CAS  CrossRef  Google Scholar 

  5. Li CJ, Nanji AA, Siakotos AN, Lin RC (1997) Acetaldehyde-modified and 4-hydroxynonenal-modified proteins in the livers of rats with alcoholic liver disease. Hepatology 26:650–657

    CAS  CrossRef  Google Scholar 

  6. Patel VB, Spencer CH, Young TA, Lively MO, Cunningham CC (2007) Effects of 4-hydroxynonenal on mitochondrial 3-hydroxy-3-methylglutaryl (HMG-CoA) synthase. Free Radic Biol Med 43:1499–1507

    CAS  CrossRef  Google Scholar 

  7. Awasthi YC, Yang Y, Tiwari NK, Patrick B, Sharma A, Li J, Awashi S (2004) Regulation of 4-hydroxynonenal-mediated signaling by glutathione S-transferases. Free Radic Biol Med 37:607–619

    CAS  CrossRef  Google Scholar 

  8. Weber D, Milkovic L, Bennett SJ, Griffiths HR, Zarkovic N, Grune T (2013) Measurement of HNE-protein adducts in human plasma and serum by ELISA-comparison of two primary antibodies. Redox Biol 1:226–233

    CAS  CrossRef  Google Scholar 

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Correspondence to Vinood B. Patel .

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Mehta, K., Patel, V.B. (2019). Measurement of 4-Hydroxynonenal (4-HNE) Protein Adducts by ELISA. In: Hancock, J., Conway, M. (eds) Redox-Mediated Signal Transduction. Methods in Molecular Biology, vol 1990. Humana, New York, NY.

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9461-8

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