14,15 EET / DHET ELISA Assay Kit

$330.00$2,620.00

The Eagle Biosciences 14,15 EET / DHET ELISA Assay kit is intended for the quantitative determination of 14,15 EET / DHET in biological samples by enzyme linked immunoassay (ELISA).  The Eagle Biosciences 14,15 EET / DHET ELISA Assay kit is for research use only and not to be used in diagnostic procedures.

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SKU: 14E39-K01 0 Categories: , ,

14,15 EET / DHET 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

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. In human carcinoma cells, rAAV-mediated over expression of CYP2J2 resulted in a marked increase in 14,15-DHET level in cell plasma, whereas rAAV-anti2J2-mediated silence of CYP2J2 expression significantly decreased its production (1).  The Eagle Biosciences 14, 15 EET/DHET ELISA Assay kit can be used to measure EET levels in cultured cells which express sEH (1).  

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 14,15-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

Manual

Product Manual