By John N. Abelson, Melvin I. Simon, Robert C. Murphy, Frank A. Fitzpatrick
This quantity comprises 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. practise and research of cytochrome P-450 metabolites of arachidonic acid are mentioned. The isolation, purification, and assay of enzymes very important within the biosynthesis and metabolism of those lipids are incorporated
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Additional info for Arachidonate Related Lipid Mediators
Acta 992, 71 (1989). 5 L. J. Roberts II, B. J. Sweetman, and J. A. Oates, J. Biol. Chem. 253, 5305 (1978). 6 M. Kumlin and E. Granstrfm, Prostaglandins 32, 741 (1986). 36 ASSAYS  closed forms required 20 days. Furthermore, the hydrolysis process is temperature dependent, increasing progressively with higher temperature. These peculiar features are essential in developing a RIA procedure for l l-dehydro-TxB2, since a carboxyl group in position l l represents a strong antigenic determinant influencing the immune response in animals as well as the affinity and specificity of antisera.
From this stock solution, prepare further dilutions in EIA buffer down to the "working highest concentration" s p. Pradelles, J. Grassi, D. Chabardes, and N. Guiso, Anal. Chem. 61, 447 (1989). 28 ASSAYS  70 6O ,--, ~9. 50 o 67 hr 40 - 47hr 30 - 16 hr 2O 10 . 0 . . . . . . . 000 . . ° . 1/10,000 Antiserum . . . . . . . . . 1/100,000 dilution FIG. I. Titration of TxB2 antiserum. Titration was performed at room temperature as described in the text. The time for the immunological reaction are indicated in the box.
4 kV. 0 × 10 -6 torr. A small amount of calibration gas (FC 43) is introduced into the calibration gas line through the automatic solenoid valve. It is then allowed to pass into the mass spectrometer by carefully opening an in-line Nupro SS-4BRW (Nupro, Willoughby, OH) needle valve. Great care is taken to avoid contamination of the mass spectrometer source with excess FC 43. 25 mA and tuning carried out on the ion at m/z 633. The electron energy is optimized for maximum sensitivity (60-90 eV, depending on the cleanliness of the source) and the source controls are then tuned for maximum sensitivity.
Arachidonate Related Lipid Mediators by John N. Abelson, Melvin I. Simon, Robert C. Murphy, Frank A. Fitzpatrick