Abstract
The assessment of prostanoids (TXB2, 6-keto-PGF1α, PGD2 and PGE2) generated in isolated or cultured cells in vitro can be directly assessed in media without extraction mainly using immunoassays, i.e. radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), or enzyme immunoassay (EIA). However, the immunoassays have to have some fundamental requisites, such as the use of highly specific and sensitive antibodies. Importantly, the results obtained with immunoassays should be validated by comparison with gas chromatography (GC), mass spectrometry (MS), or liquid chromatography (LC)–MS. The measurement of prostanoid generation in vivo can be performed by the evaluation of prostaglandin breakdown products in urine using LC–MS, GC–MS, or validated immunoassays after extraction and rigorous sample purification. The measurement of F2-isoprostanes (F2-iPs), containing F-type ring analogous to PGF2α, provides a reliable tool for identifying populations with enhanced rates of lipid peroxidation. Levels of 8- iso-PGF2α (also referred as iPF2α-III) in plasma or urine are assessed by GC/MS or validated immunoassays.
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Patrignani, P. (2010). Extraction and Measurement of Prostanoids and Isoprostanes: Introduction to Part II. In: Ayoub, S., Flower, R., Seed, M. (eds) Cyclooxygenases. Methods in Molecular Biology, vol 644. Humana Press. https://doi.org/10.1007/978-1-59745-364-6_12
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DOI: https://doi.org/10.1007/978-1-59745-364-6_12
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