Cardiovascular and Oxidative Stress Assay Kit

11,12 DHET ELISA Assay Kit

$405.00

The 11, 12-DHET ELISA Assay kit is intended for the quantitative determination of 11, 12-DHET in biological samples by enzyme linked immunoassay (ELISA).  The Eagle Biosciences 11, 12 DHET ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

SKU: 11D39-K01 Categories: , ,

11,12 DHET ELISA Assay Kit

The 11,12 DHET ELISA Assay Kit is For Research Use Only
Product manufactured in the USA
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

Assay Background

The level of 11, 12-DHET or 11, 12-DHET epitope has been shown to exhibit correlation with hypertension in rodents (1, 2, 3).  11, 12-DHET is a representative metabolite of cytosolic epoxide hydrolase-mediated metabolism of EETs, which are generated by arachidonic acid epoxygenase activity of cytochrome P450’s (4).  The 11,12-DHET kit can be used for the determination of 11,12-DHET 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.

The competitive 11,12-DHET ELISA, based on competition between 11,12-DHET epitope and 11,12-DHET-HRP conjugate for a limited number of binding sites available from the anti-11,12-DHET antibody, which is coated to the wells of the 96 well ELISA plate.  The conjugate concentration is held as a constant in each well, while the concentration of the 11,12-DHET is variable, based on the concentration of the sample or standard.  Thus the amount of the 11,12-DHET conjugate which is able to bind to each of the wells is inversely proportional to the concentration of 11,12-DHET in the standard or sample.  The amount of the conjugate which is bound to each well is then determined by the amount of color obtained, when TMB is added.  The TMB reacts with the HRP available in the well.  With the addition of sulfuric acid, the blue colored product is converted into a yellow colored product, which can be read on a plate reader at 450 nm.

Related Products

14,15 DHET ELISA Assay Kit
20-HETE ELISA Assay Kit
14,15 EET DHET ELISA Assay Kit

Additional Information

Assay Procedure

  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 11,12-DHET-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.

Package Inserts

 

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.