Linoleic acid, the predominant polyunsaturated fatty acid (PUFA) in the human diet, can be metabolized by cyclooxygenase, lipoxygenase and P450 enzymes. The hydroxyoctadecadienoic acid (HODE) derivatives of linoleic acid, 9(R)-HODE, 9(S)-HODE and 13(S)-HODE, are the most widely distributed of the known linoleic acid metabolites.
These compounds exhibit interesting biologic activities, including the regulation of platelet function, maintenance of vascular thromboresistance and transduction of the cellular responses to certain growth factors. HODE derivatives may also influence certain pathological states including, psoriasis, the development of atherosclerosis and the development of cancer.
Principle
This assay measures the level of total 9-HODE, which includes both 9(S)-HODE and 9(R)-HODE, in biological samples. This kit is a competitive ELISA. Briefly, the 9-HODE present in the samples or standards competes with 9(±)-HODE conjugated to HRP [9(±)-HODE-HRP] for binding to an antibody specific for 9(±)-HODE that is precoated on a microplate. The peroxidase activity of 9(±)-HODE-HRP results in color development when a substrate is added. The intensity of the color is proportional to the amount of 9(±)-HODE-HRP bound and is inversely proportional to the amount of unconjugated 9-HODE present in the samples or standards.
Kit Components
H9110-18A1: Microtiter Plate, 1x96 wells H9110-18A2: Standard, 1x20ul, 0.05mg/ml in ethanol H9110-18A3: Wash Buffer, 5X, 1x50ml H9110-18A4: Dilution Buffer, 5X, 1x25ml H9110-18A5: TMB Substrate, 1x25ml H9110-18A6: 9(±)-HODE-HRP conjugate, 1x320ul
Storage and Stability
Store components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.