| Product name | Human 5-hydroxyeicosatetraenoic acid (5-HETE) ELISA Kit |
| Reactivity | Human |
| Applications | ELISA |
| Applications notes | This Human 5-hydroxyeicosatetraenoic acid (5-HETE) ELISA Kit employs a two-site sandwich ELISA to quantitate 5-HETE in samples. An antibody specific for 5-HETE has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any5-HETE present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for 5-HETE is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of 5-HETE bound in the initial step. The color development is stopped and the intensity of the color is measured. |
| Detection method | Colorimetric |
| SampleType | Cell culture supernatants, Other biological fluids, Plasma, Serum |
| Assay type | Sandwich ELISA (quantitative) |
| Assay duration | Multiple steps standard sandwich ELISA assay with a working time of 3-5 hours. It depends on the experience of the operation person. |
| Alternative | 5-HETE |
| Kit components | • Human 5-hydroxyeicosatetraenoic acid microplate • Human 5-hydroxyeicosatetraenoic acid standard • Human 5-hydroxyeicosatetraenoic acid detect antibody • Streptavidin-HRP • Standard diluent • Assay buffer • HRP substrate • Stop solution • Wash buffer • Plate covers |
| Features & Benefits | Human 5-hydroxyeicosatetraenoic acid (5-HETE) ELISA Kit has high sensitivity and excellent specificity for detection of Human 5-HETE. No significant cross-reactivity or interference between Human 5-HETE and analogues was observed. |
| Calibration range | Please inquire |
| Limit of detection | Please inquire |
| Usage notes | • Do not mix components from different kit lots or use reagents beyond the kit expiration date. • Allow all reagents to warm to room temperature for at least 30 minutes before opening. • Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination. • Unused wells must be kept desiccated at 4 °C in the sealed bag provided. • Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes. • It is recommended that all samples and standards be assayed in duplicate or triplicate. |
| Storage instructions | The unopened kit should be stored at 2 - 8°C. After opening, please store refer to protocols. |
| Shipping | Gel pack with blue ice. |
| Precautions | The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product. |
| Background | 5-Hydroxyeicosatetraenoic acid (5-HETE) is an endogenous eicosanoid. 5-HETE is an intermediate in the pathway of leukotriene synthesis. In addition, it is a modulator of tubuloglomerular feedback. 5-Hydroxyeicosatetraenoic acid is a key intermediate of the arachidonate-dependent protective signaling in monocytes/macrophages exposed to peroxynitrite. Peroxynitrite elicited the nuclear membrane translocation of 5-LO and enhanced its enzymatic activity via a mechanism sensitive to low concentrations of inhibitors of 5-LO or the 5-LO-activating protein, as well as to genetic depletion of the latter enzyme. Inhibition of 5-LO activity was invariably associated with the cytosolic localization of PKC, the mitochondrial accumulation of Bad, and a rapid MPT-dependent necrosis. All these events were prevented by nanomolar concentrations of the 5-LO product 5-hydroxyeicosatetraenoic acid. |
| Alternative | 5-HETE |
Fig.1. Human 5-hydroxyeicosatetraenoic acid (5-HETE) Standard Curve.
Fig.2. Abbkine ELISA kit is series of sandwich ELISA to quantitate specific protein in samples.
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1.The species of antibody reactivity should be the sample species that can be matched normally after Abbkine R&D experts have passed strict scientific verification. If your sample is not within the range of reactivity, in order to improve the efficiency and results of your experiment, it is not suggested to try other species. Otherwise, it may lead to sample mismatch and affect the effect of your experiment.
2.Please aliquot the antibody received as soon as possible and store it at -20℃, avoid repeated freezing and thawing, and use it within one year.
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