By John N. Abelson, Melvin I. Simon, Robert C. Murphy, Frank A. Fitzpatrick
This quantity contains details on arachidonic acid metabolites and platelet activating issue. Assays for the quantitative dimension of prostaglandins, leukotrienes, and platelet-activating issue, together with enzyme immunoassays and mass spectrophotometric options, are offered. coaching and research of cytochrome P-450 metabolites of arachidonic acid are mentioned. The isolation, purification, and assay of enzymes vital within the biosynthesis and metabolism of those lipids are integrated
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Additional info for Arachidonate Related Lipid Mediators
Patrono, G. Ciabattoni, and G. A. FitzGerald, Anal. Biochem. 155, 198 (1986).  RADIOIMMUNOASSAY OF 11-DEHYDROTHROMBOXANEB2 37 90% of lactone form into the open form. The hydrolysis reaction probably occurs within the dermis of the immunized animal because (a) being emulsified in Freund's adjuvant, the conjugate is slowly adsorbed within several days and (b) body temperature strongly increases the conversion rate. This particular behavior of l l-dehydro-TxB2 has two main consequences: (a) hydrolysis unmasks a strong antigenic determinant in a strategic position and the molecule attached through the C-1 terminus protrudes out of the carrier protein, thus leaving the C-11 carboxyl exposed to macrophage recognition; (b) antibody specificity will be mainly directed against this portion of the molecule, which is unique among the metabolites produced by cyclooxygenation of arachidonate.
T ......... 200 300 400 m/z FIG. 5. NCI mass spectra of (a) 14,15-EET PFB ester; (b) H6-14,15-EET PFB ester. This methodology has facilitated a number of studies on the role that lipid mediators play in both normal and pathophysiological processes. Method for Quantitative Analysis of 6-Oxo-PGFI,~ by Capillary GC-NCI-MS General Procedure HPLC-grade solvents (B & J , American Scientific, McGaw Park, IL) were used throughout the study. Pyridine (HPLC grade) was obtained from Aldrich Chemical Co. (Milwaukee, WI).
300 , ......... 400 m/z , ......... 500 ~ ,A ....... 600 I 700 1614 I c~, >~ / o/ ....... ~ . . . . . , ......... , ......... , ....... I ,oo FIG. 4. HCI mass spectrum of (a) MO, Tris-TMS ether, PFB ester derivative of 6-oxoPGF1~; (b) MO, Tris-TMS ether, PFB ester derivative of TxB2. on phenyl boronate minicolumns, 25 immunoaffinity chromatography, 26'27 and tandem mass spectrometry. 2s The excellent resolution that can be obtained with capillary GC has encouraged investigators to develop assays that allow quantification of several mediators in a single chromatography run.
Arachidonate Related Lipid Mediators by John N. Abelson, Melvin I. Simon, Robert C. Murphy, Frank A. Fitzpatrick
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