Abstract
A radioimmunoassay (RIA) for human pro-LRF (14-69) OH was developed with an antiserum, generated in a rabbit, to [Tyr67] pro-LRF (47-67) NH2 conjugated to BSA. This antiserum bound 28-32% of [125I] pro-LRF (14-69) 0H at a final dilution of 1:2500 and the binding was inhibited by pro-LRF (14-69) OH in a dose-dependent manner. The sensitivity of the RIA was 31.2-62.5 pg and the dose that inhibited 50% of the binding to the tracer was 280-320 pg. Intra-and inter-assay coefficients of variation at 50% inhibition were 8 and 12%, respectively. Neither LRF nor pro-LRF (14-37) OH was recognized by the antiserum. The dilution curve generated with human hypothalamic extract was parallel to that of pro-LRF (14-69) OH. In addition the extract yielded a major immunoreactive peak emerging in elution volumes concordant with [125I] pro-LRF (14-69) OH on Sephadex G-SO chromatography.
