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The multispecies Prostaglandin E2 (PGE2) ELISA quantitates PGE2 in serum, plasma, and other biological fluids
原理方法:
The Competitive multispecies ELISA research-use-only kit is designed to quantitatively measure Prostaglandin E2 (PGE2) independent of species. A PGE2 standard is provided to generate a standard curve for the assay and all samples are read off a user-generated standard curve. Standards or diluted samples are pipetted into a coated microtiter plate and Biotinylated Detection Ab specific to PGE2 is added. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured on a microplate reader. The concentration of PGE2 in the samples is then determined by comparing the OD of the samples to the standard curve.
Rigorous validation:
Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.
Prostoglandin E2 (PGE2) is a potent lipid mediator produced by metabolism of arachidonic acid via the cyclooxygenase (COX) pathway. Following cellular stimulation, arachidonic acid is hydrolyzed from phospholipids stores by one of a family of phospholipase A2 enzymes and converted to the unstable endoperoxide PGH2 by either COX 1 or COX 2. PGH2 is isomerized to PGE2 by one of 3 distinct enzymes microsomal PGES 1 (mPGES 1), mPGES 2, or a cytosolic enzyme, cPGES. PGE2 is active in a number of physiological systems and settings including inflammation, immune regulation, generation of fever, pain perception, protection of the gastric muscosa, fertility and parturition, as well as sodium and water retention. The effects of PGE2 are transduced by 4 subtypes of G protein coupled receptors designated EP1, EP2, EP3, EP4, with Kd values for PGE2 ranging from 1 10 nM. PGE2 is rapidly metabolized in vivo by the prostaglandin 15 dehydrogenase pathway (15 hydroxy PGDH) to the inactive metabolite 13,14 dihydro 15 keto PGE2. The half life of PGE2 in the circulatory system is ~30 seconds and normal plasma levels are 3 12 pg/mL
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