A radioimmunoassay for leukotriene B4 |
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Authors: | John A Salmon Paula M Simmons Richard MJ Palmer |
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Institution: | Depts. of Prostaglandin Research and Pharmacology, Welcome Research Laboratories, Langley Court, Beckenham, Kent BR3 3BS, UK |
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Abstract: | A radioimmunoassay for leukotreine B4 has been developed. The assay is sensitive; 5 pg LTB4 caused significant inibition of binding of 3H]-LTB4 and 50% displacement occurred with 30 pg. The specificity of the assay has been critically examined; prostaglandins, thromboxane B2 and arachidonic acid do not exhibit detectable cross-reactions (< 0.03%). Flowever, some non-cyclic dihydroxy- and monohydroxy-eicosatetraeonic acids do cross-react slightly (e.g. diastereomers of 5,12-dihydroxy-6,8,10--14--elcosatetraenoic and 12-hydroxy-5,8,10,14-elcosatetraenoic acids cross-react 3.3% and 2.0% respectively). The assay has been used to monitor the release of LTB4 from human neutrophils in response to the divalent cation ionophore, A23187. The immunoreactive material released during these incubations was confirmed as LTB4 by reverse-phase high liquid chromatography follwing solvent extraction and silinic acid chromatography. |
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