20-HETE ELISA Assay Kit

$330.00$2,620.00

The Eagle Biosciences 20-Hete ELISA Assay kit is intended for the quantitative determination of 20-Hete (20-hydroxyeicosatetraenoic acid) in biological samples by enzyme linked immunoassay (ELISA). The 20-Hete ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

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SKU: 20H39-K01 0 Categories: , ,

20-HETE ELISA Assay Kit

For Research Use Only

Size: 1×96 wells
Sensitivity: 0.01 ng/ml
Dynamic Range: 0.01 – 1000 ng/ml
Incubation Time: 2.5 hours
Sample Type: Serum, Plasma, Tissue, Biological Fluids
Sample Size: 100 µl

Product manufactured in the USA

Additional Information

Assay Background


The specificity of the 20-HETE ELISA Assay kit was investigated using authentic 20-HETE and a panel of fatty acids which, based on their structure, might be anticipated to compete with 20-HETE for binding to antibodies for 20-HETE.  Anti-20-HETE did not cross-react with 14, 15- and 11,12-DHETs, PGE2 and showed almost no cross-reactivity even with structurally extremely similar arachidonic acid (AA), linoleic acid and linolenic acid as shown in the competitive ELISA analysis.  Considering the only difference between 20-HETE and AA is an oxygen molecule, the specificity of the Eagle Biosciences 20-HETE ELISA is a surprise.

Human essential and salt-sensitive hypertensions were related to differential AA metabolism by cytochrome P450 (CYP) 4A which has AA-ω-hydroxylase (20-HETE synthesis) activity.  Increased circulating insulin inhibits 20-HETE synthesis in obese hypertensive subjects.

Recently, CYP4F2 genetic variants, which increased urinary 20-HETE secretion, were found to be correlated with the risk for hypertension in a Chinese population.   The 20-Hete ELISA Assay kit can be used for the determination of 20-HETE in serum, plasma, cells, and tissues following proper isolation and purification.  Instructions are provided as to the proper isolation and purification in the following pages.

Assay Principle


  1. Load 200 microliters of Sample Dilution Buffer into the blank (BL) wells and 100 microliters of Sample Dilution Buffer into the maximum binding (BO) wells.
  2. Load 100 microliters of each of the standards into the appropriate wells.
  3. Load 100 microliters of each of the samples into the appropriate wells.
  4. Load 100 microliters of the diluted 20-HETE-HRP conjugate in the BO wells, the standard wells, and the sample wells.  Do NOT add HRP conjugate into the BL wells.
  5. Incubate the plate at room temperature for two hours.
  6. Wash the plate three times with 400 microliters of the diluted Wash Buffer per well.
  7. After the last of the three wash cycles pat the plate dry onto some paper toweling.
  8. Add 200 microliters of the TMB substrate to all of the wells (including BL wells).
  9. Incubate the plate at room temperature for 15-30 minutes.
  10. Add 50 micoliters of 2 N sulfuric acid to all of the wells.
  11. Read the plate at 450 nm.

Manual

Product Manual


Publications

References


  • Liu, H., Zhao,Y., Nie, D., Shi, J., Fu, L., Li, Y., Yu, D., and Lu, J. Association of a Functional Cytochrome P450 4F2 Haplotype with Urinary 20-HETE and Hypertension. J Am Soc Nephrol 19: 714-721, 2008
  • Meseguer et al. Kidney Androgen-Regulated Protein Transgenic Mice Show Hypertension and Renal Alterations Mediated by Oxidative Stress.  Circulation 119, 1908-1917, 2009.
  • Domaski et al.  Is it possible to predict the early post-transplant allograft function using 20-HETE measurements? A preliminary report. Transplant International 22, 546-553, 2009
  • Liu et al.  Overexpression of cytochrome P450 4F2 in mice increases 20-hydroxyeicosatetraenoic acid production and arterial blood pressure.  Kidney International 75, 1288-1296, 2009.
  • Wang et al.  Selective Inhibitors of CYP2J2 Related to Terfenadine Exhibit Activity Strongly against Human Cancers in vitro and in vivo.  JPET, 152017, 2009.

Recent Citations


Yi, M;Cho, SA;Min, J;Kim, DH;Shin, JG;Lee, SJ;, (2017). Functional characterization of a common CYP4F11 genetic variant and identification of functionally defective CYP4F11 variants in erythromycin metabolism and 20-HETE synthesis. Arch. Biochem. Biophys. PubMed: 28347661