Background

Prostaglandin E2 (PGE2) is derived from Prostaglandin H2 (PGH2), which in turn is synthesized from arachidonic acid, through the cyclooxygenase pathway. Many cell types, such as, epithelial cells, fibroblasts, and macrophages produce PGE2. PGE2 possesses vasoactivity, modulates immune functions, regulates sleep-wake cycle and exhibits many other activities. This competitive ELISA kit provides quantitative analysis of PGE2 levels in biological fluids based on the competition between the PGE2 present in the sample and PGE2-enzyme conjugate for a limited number of antibody binding sites.

Assay Principle

This more specific ELISA for the quantitative analysis of PGE2 levels in biological fluid operates on the basis of competition between the enzyme conjugate and the PGE2 in the sample for a limited number of binding sites on the antibody-coated plate. The sample or standard solution is first added to the microplate. Next, the diluted enzyme conjugate is added and the mixture is shaken and incubated at room temperature for one hour. During the incubation, competition for binding sites is taking place. The plate is then washed removing all the unbound material. The bound enzyme conjugate is detected by the addition of substrate which generates an optimal color after 30 minutes. Quantitative test results may be obtained by measuring and comparing the absorbance reading of the wells of the samples against the standards with a microplate reader at 450 or 650nm. The extent of color development is inversely proportional to the amount of PGE2 in the sample or standard. For example, the absence of PGE2 in the sample will result in a bright blue color, whereas the presence of PGE2 will result in decreased or no color development.

If using urine as a sample it is recommended to run our

Creatinine Assay kit (CR01)

in conjunction with your samples since urinary creatinine levels may be used to normalize the rate of excretion of other analytes.