Description
Key facts
Detection method:Colorimetric,
Sample types:Purified protein, Tissue Lysate, Plasma, Serum, Cell Lysate,
Assay type:Competitive,
Assay Platform:Microplate reader
Product details:
How the assay works
This Lipid Peroxidation (4-HNE) Assay Kit will allow the quantitation of 4-HNE adduct in protein samples determined by comparing its absorbance with that of a known 4-HNE-BSA standard curve.4-HNE is a well known by-product of lipid peroxidation and is widely accepted as a stable marker for oxidative stress.
Lipid Peroxidation (4-HNE) Assay Kit protocol summary:
- Prepare all reagents, samples, and standards as instructed.
- Add 50 µL standard or sample to wells of 4-HNE Conjugate coated plate and incubate for 10 mins. Add 50 µL of the diluted anti-4-HNE antibody and incubate for 1 h.
- Washing steps with 250 µL 1X Wash Buffer.
- Add 100 µL diluted Secondary Antibody-HRP Conjugate per well and incubate for 1 h. Wash as before with 1X Wash buffer.
- Add 100 µL of warm Substrate Solution and incubate for 2-20 mins.
- Stop the enzyme reaction by adding 100 µL of Stop Solution to each well. Read absorbance immediately on a microplate reader using 450 nm.
How other researchers are using
Lipid Peroxidation (4-HNE) Assay Kit has been used in variety of sample type including:
- Mouse osteoclast cells 1
- Rat brain tissues 2
- Human glioma tissues 3
References:
1 - Louka P et al. 2024; 2 - Davaanyam D et at 2023; 3 - Wang Z et al. 2023.
Related and recommended products
See below alternatives Lipid Peroxidation Assay Kits:
- Lipid Peroxidation Assay Kit (Cell-based)
ab243377
- Lipid Peroxidation (MDA) Assay Kit (Colorimetric/Fluorometric)
ab118970
- Lipid Peroxidation (MDA) Assay Kit (Colorimetric)
ab233471
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
4-Hydroxynonenal (4-HNE) often referred to as HNE is a highly reactive aldehyde with a molar mass of approximately 156.19 g/mol. It plays a significant role in lipid peroxidation processes. 4-HNE forms during the oxidative degradation of polyunsaturated fatty acids and is present mostly in tissues with high metabolic activity. It is frequently expressed in the liver brain and heart where oxidative stress is elevated. Researchers commonly use techniques like 4-HNE staining and 4-HNE Western blot to study its presence and implications in various tissues and cells.
Biological function summary
4-HNE serves as a signaling molecule influencing many cellular pathways. It is not just a byproduct of oxidative stress but actively modifies proteins through covalent binding. These modifications often lead to changes in protein function and structure affecting cellular processes such as proliferation apoptosis and differentiation. 4-HNE also interacts with other reactive molecules and antioxidants forming a complex network of cellular responses. This aldehyde regulates gene expression influencing how cells respond to oxidative stress and inflammation.
Pathways
4-HNE interacts with pathways involved in oxidative stress and detoxification. In the oxidative stress pathway it reacts with glutathione to form glutathione conjugates which are part of the cell’s detoxifying responses. The aldehyde thereby affects the Keap1-Nrf2-ARE pathway which modulates antioxidant responses and cellular protection against oxidative damage. Proteins like glutathione peroxidase and glutathione-S-transferase play key roles in these pathways illustrating the interconnectedness of 4-HNE with cellular defense mechanisms.
4-HNE has associations with conditions such as Alzheimer's disease and cardiovascular diseases. In Alzheimer's the aldehyde contributes to neuroinflammation and protein modification impacting neuronal function and survival. It interacts with proteins like tau and amyloid-beta which are critical in the pathology of Alzheimer's. In cardiovascular diseases 4-HNE mediates cellular damage and inflammation linking it to the progression of atherosclerosis. This reactive aldehyde interacts with proteins such as apolipoprotein B highlighting its significant role in disease pathways linked to oxidative stress.
