Use of tritium labeled amino acid conjugates of prostaglandins and thromboxanes as labeled ligands in prostanoid radioimmunoassay.
Conjugates of prostaglandins and thromboxanes with tritium labeled amino acids were prepared and employed as labeled ligands in prostaglandin and thromboxane radioimmunoassays. Assays for PGF2 alpha, 15-keto-13, 14-dihydro-PGF2 alpha, TXB2 and 15-keto-13,14-dihydro-TXB2 were evaluated in comparative studies using either these heterologous ligands or the corresponding homologous tritiated eicosanoid as tracers. Binding properties for the respective antibodies were found to be similar using either tracer. Three biological studies were also conducted, viz. study of the release of TXB2 during collagen induced platelet aggregation, of 15-keto-13,14-dihydro-TXB2 during guinea pig pulmonary anaphylaxis, and of PGF2 alpha (measured as 15-keto-13,14-dihydro-PGF2 alpha in peripheral plasma) during bovine luteolysis. The analyses gave comparable results using either the heterologous or the homologous assay. Thus, this type of labeled prostanoid conjugates may serve as a convenient alternative to homologous tracers in radioimmunoassay. Heterologous tracers may even in certain cases provide the only simple solution to the problem of preparing a labeled ligand of high specific activity.
D000596:Amino Acids; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D008024:Ligands; D011453:Prostaglandins / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods*; D013931:Thromboxanes / Q000032:analysis*; D014316:Tritium
D000596:Amino Acids; D008024:Ligands; D011453:Prostaglandins; D013931:Thromboxanes; D014316:Tritium