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4-Hydroxynonenal ELISA Kit (4-HNE)

Full Name: 4-Hydroxynonenal ELISA Kit (4-HNE)
Reactivity: Universal
Sample Type: Tissue Homogenates, Plasma, Serum, Biological Fluids
Sensitivity: 18.75 pg/ml

INTRODUCTION

4-Hydroxynonenal (4-HNE) is formed through a process of lipid peroxidation which leads to production of an alpha, beta unsaturated. It can be described as a colourless oil and it is present in majority of animal tissues. The levels present are elevated during periods of increased levels of stress. One of the key functions of is during various cell signal transduction pathways, that can include cellular adhesion and cell cycle events.

4-HNE is a degradation product of the fatty acid peroxidation product malondialdehyde. In general, it reacts with reduced glutathione catalyzed by Glutathione peroxidase. In some cases, it has been found to react with cysteine under oxidative stress conditions and this reaction leads to formation of mercapturic acid thiol which then reacts further with DNA resulting in cytotoxicity.

4-Hydroxynonenal has been found to have a hormetic and dual action on the health of cells. Presence of lower intracellular concentrations are known to benefit cells by promoting proliferation, antioxidant defence, differentiation, and other compensatory mechanisms. Whilst higher concentrations of 4-HNE are known to activate various toxic pathways with the eventual outcome of cell death either through a process of necrosis or apoptosis. Examples of this include laddering of genomic DNA, induction of caspase enzymes and the release of mitochondrial cytochrome c.

INTENDED USE

4-Hydroxynonenal ELISA kit can be used for detecting levels of 4-HNE (4-hydroxynonenal) using samples of plasma, serum, other biological fluids and tissue homogenates.

 CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

  • One 96-Well Plate: Pre-coated with anti 4-HN4 antibody.
  • Standards: Lyophilized recombinant.
  • Sample/Standard Dilution Buffer.
  • Biotinylated-labelled Antibody.
  • Antibody Dilution Buffer.
  • HRP-Streptavidin Conjugate (SABC).
  • SABC Dilution Buffer.
  • TMB Substrate.
  • Wash Buffer (25x).
  • Plate Sealer.
  • Product Instructions.

TYPICAL RESULTS

For the following 4-Hydroxynonenal ELISA kit  it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 31.25, 62.5, 125, 250, 500, 1000, 2000 pg/ml.
Reactivity: Universal
Sensitivity: 18.75 pg/ml
Range: 31.25 – 2000 pg/ml
Principle: Competitive
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for 4-HNE, no cross reactivity or interference between 4-hydroxynonenal and analogues was detected.
– Recovery: Serum (85 – 102%), EDTA Plasma (88 – 97%), Heparin Plasma (86 – 104%).
– Linearity: Serum (86 – 101%), EDTA Plasma (81 – 100%), Heparin Plasma (91 – 101%).
– Precison Intra-Assay: CV < 8%.
– Precison Inter-Assay: CV < 10%.
– Stability: Less than 10%.

REFERENCES

  1. 4-hydroxynonenal-mediated signaling and aging. Free Radic Biol Med. (2017) 111: 219-225. Zhang H. and Forman H.J.
  2. Salivary Redox Biomarkers in Insulin Resistance: Preclinical Studies in an Animal Model. Oxid Med Cell Longev. (2021) 2021: 3734252. Maciejczyk M., et al.
  3. 4-HNE Immunohistochemistry and Image Analysis for Detection of Lipid 4.
  4. Peroxidation in Human Liver Samples Using Vitamin E Treatment in NAFLD as a Proof of Concept. J Histochem Cytochem. (2020) 68 (9): 635-643. Podszun M.C., et al.

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