Competitive enzyme immunoassays (EIA) are employed for the quantitative determination of prostaglandin E1 (PGE1) and prostaglandin E2 (PGE2) in biological fluids. In the assays, the specific protein in the sample competes with a fixed amount of alkaline phosphatase-labeled conjugate for binding sites on the antibody. The intensity of the bound yellow color read at 405 nm is inversely proportional to the concentration of prostanoid in the standards or samples. The measured optical density is used to calculate the concentration.
Biological actions include vasodilation, both anti-and pro-inflammatory action, modulation of sleep/wake cycle, stimulation of bone resorption and thermoregulation.
Prostaglandins and thromboxane are products of the cyclooxygenase pathway and are synthesized in response to immediate need and are not stored in significant amounts for later release. Prostanoids mediate various biological functions in humans, including platelet aggregation, gastric mucosa protection, renal homeostasis, vascular homeostasis, uterine function, embryo implantation and labor and sleep/wake cycle.
Features and Benefits
The assays specifically measure each target lipid with negligible cross-reactivity with 20-25 related eicosanoid compounds tested (from <0.1 to 2%). A standard curve is run in each assay, precoated plates eliminate the coating step, all incubations proceed at room temperature, any commercially available microwell plate reader may be used and reagents are color coded for easy use. The results are available in 4 hours. Performance characteristics show low intra- and inter-assay variability, linear standard curve and recovery of samples ranging from 99-111%.
The sensitivity of this EIA is 15.9 pg/mL and it specifically detects human PGE2 in cell culture supernatants, saliva, urine, plasma and serum without interference from other prostanoids.