14,15 EET / DHET ELISA Kit

Product details

Abcam's 14,15 EET / DHET in vitro ELISA kit is designed for accurate quantitative measurement in serum, plasma, urine, cell culture, tissue samples following proper isolation and purification.

A 96-well plate has been precoated with 14,15-DHET antibody. Samples and the 14,15-DHET-HRP conjugate are added to the wells, where 14,15-DHETin the sample competes with the added 14,15‑DHET-HRP for antibody binding. After incubation, the wells are washed to remove unbound material and TMB substrate is then added which is catalyzed by HRP to produce blue coloration. The reaction is terminated by addition of Stop Solution which stops the color development and produces a color change from blue to yellow. The intensity of signal is inversely proportional to the amount of 14,15‑DHET in the sample and the intensity is measured at 450 nm.

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. Abcam's14,15EET/DHET in vitro ELISA kit can be used to measure EET levels in cultured cells which express sEH.