4-Hydroxynonenal (4-HNE),a prominent lipid peroxidation product, is a highly reactive aldehyde generated during the oxidative degradation of polyunsaturated fatty acids in cell membranes. This molecule has garnered significant attention due to its profound influence on a wide range of biological and pathological processes. It exerts its effects by forming covalent adducts with proteins, nucleic acids, and lipids, thereby altering their structure and function.

Extensive research demonstrates that 4-HNE is a critical mediator in the pathogenesis of various diseases, including cancer, neurodegenerative disorders, cardiovascular diseases, and metabolic syndromes. Moreover, 4-HNE plays a pivotal role in modulating key cellular signaling pathways, such as oxidative stress response, inflammation, and apoptosis. These interactions highlight its dual nature as both a biomarker and a modulator of disease development and progression.

The RayBio^® 4-HNE ELISA Kit is a competitive enzyme immunoassay developed for rapid detection and quantitation of 4-HNE in serum, plasma, cell supernatant or urine samples collected from various species including humans, rats or mice. The quantity of 4-HNE is determined by a known 4-HNE standard curve. This kit has a high sensitivity of approximately 0.363 ng/mL. Each kit provides sufficient reagents to perform up to 96 assays, including standard curve and unknown samples.

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

• Microplate reader capable of measuring absorbance at 450 nm
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1-liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• GraphPad Prism or SigmaPlot software (or other software which can perform four-parameter logistic regression models)
• Tubes to prepare standard or sample dilutions
• Orbital shaker
• Aluminum foil
• Plastic wrap

1. Prepare all reagents, samples and standards as instructed.
2. Add 50 μL standards or samples to appropriate well. Incubate for 10 minutes at room temperature.
3. Add 50 μL anti-4-HNE to appropriate well. Incubate 1 hour at room temperature. And wash 5 times.
4. Add 100 μL prepared HRP-Secondary antibody solution. Incubate 1 hour at room temperature. And wash 5 times.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 2-10 minutes at room temperature.
6. Add 50 μL Stop Solution to each well. Read at 450 nm immediately.

The entire kit may be stored at -20°C to -80°C for up to 1 year from the date of shipment. For extended storage, it is recommended to store it at -80°C. Avoid repeated freeze-thaw cycles.