Eskra J D, Levine L, Carty T J
Prostaglandins Med. 1980 Sep;5(3):201-7.
An efficient method for the enzymic preparation of high specific radioactivity [3H]-12-L-hydroxyeicosatetraenoic acid (12-L-HETE) is described. This compound was used as a radiolabeled ligand in the radioimmunoassay (RIA) of 12-L-HETE. The accuracy of the RIA was checked by incubating [14C]arachidonic acid with platelet lipoxygenase, and measuring enzyme activity in the presence of the inhibitor, 1-phenyl-3-pyrazolidinone (phenidone). The amount of 12-L-HETE synthesized, determined by RIA, was found to be in agreement with that analyzed by radiochemical assay after thin layer plate chromatography.
描述了一种酶促制备高比放射性[3H]-12-L-羟基二十碳四烯酸（12-L-HETE）的有效方法。该化合物用作12-L-HETE放射免疫分析（RIA）中的放射性标记配体。通过将[14C]花生四烯酸与血小板脂氧合酶一起孵育，并在抑制剂1-苯基-3-吡唑烷酮（非那宗）存在下测量酶活性，来检查RIA的准确性。通过RIA测定合成的12-L-HETE量，发现与薄层板色谱后放射化学分析的结果一致。
