Description
Principle of the assay: This assay employs the competitive enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with goat-anti-rabbit antibody. Standards or samples are added to the appropriate microtiter plate wells with an antibody specific for PG-E2 and Horseradish Peroxidase (HRP) conjugated PG-E2. The competitive inhibition reaction is launched between with HRP labeled PG-E2 and unlabeled PG-E2 with the antibody. A substrate solution is added to the wells and the color develops in opposite to the amount of PG-E2 in the sample. The color development is stopped and the intensity of the color is measured.
Background: Prostaglandin E2 (PG-E2) is a primary product of arachidonic acid metabolism in many cells. Like most eicosanoids, it does not exist preformed in any cellular reservoir. When cells are activated or exogenous free arachidonate is supplied, PG-E2 is synthesized de novo and released into the extracellular space. In vivo, PG-E2 is rapidly converted to an inactive metabolite (13,14-dihydro-15-keto PG-E2) by the prostaglandin 15-dehydrogenase pathway. The half-life of PG-E2 in the circulatory system is approximately 30 seconds and normal plasma levels are 3-12 pg/ml. In general, urine and culture media samples can be diluted, if necessary, and added directly to the assay well. Plasma samples should be purified prior to use. Because of the rapid metabolism of PG-E2, the determination of in vivo PG-E2 biosynthesis is often best accomplished by the measurement of PG-E2 metabolites