Cardiovascular and Oxidative Stress Assay Kit

11,12 EET DHET ELISA Assay Kit

$425.00

The 11,12 EET 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 EET/DHET ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

SKU: 11E39-K01 Categories: , ,

11,12 EET DHET ELISA Assay Kit

The 11,12 EET 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

Storage and Stability
The 11,12-EET/DHET Assay kit will obtain optimal results if all of the components are stored at the proper temperature prior to use. Items should be stored at the designated temperatures upon receipt of the 11,12-EET/DHET ELISA. All components are stored below -20°C and should not be re-frozen and thawed more than necessary.
Sample Preparation
EET+DHET can be measured after chemically changing EET to DHET. However, if the EET in cells or in blood is changed to DHET by abundantly expressed soluble epoxide hydrolase, measurement of DHET without chemically changing EET to DHET is suitable.
However, when P450 2C23 activity of the rat microsomes was measured, the rat microsomes were incubated with arachidonic acid (substrate of P450 2C23) and, then, EET + DHET levels in the reaction mixture were measured after acid hydrolysis of EET to DHET, which was indicative of P450 2C23 activity.
There are three different protocols which can be used to convert EET into DHET for measurement using the competitive ELISA kit. For optimal results please choose the protocol which fits your sample best.

Assay Background

It is well known that arachidonic acid (AA) will be converted to EET by P450 arachidonic acid epoxygenase (AA epoxygenase) and EET will be converted to DHET by soluble epoxide hydrolase (sEH) in vivo.  Cytochrome P450 2J2 (CYP2J2) is a predominant human AA epoxygenase that produces all four EETs. The EET/DHET ELISA Assay kit can be used to measure EET levels in cultured cells which express sEH (1).

Related Products

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

Typical Standard Curve

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.

Product Citations