A radioimmunoassay for human pro-luteinizing hormone-releasing factor [pro-LRF(14-69)OH].
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)OH 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-50 chromatography.
Motoyama
A
A
Wakabayashi
I
I
Minami
S
S
Sugihara
H
H
Takahashi
F
F
Akira
S
S
Ling
N
N
D000918:Antibody Specificity; D007987:Gonadotropin-Releasing Hormone / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D011498:Protein Precursors / Q000032:analysis* / Q000276:immunology; D011786:Quality Control; D011863:Radioimmunoassay
D007106:Immune Sera; D011498:Protein Precursors; C053348:pro-LHRH (14-69)OH; D007987:Gonadotropin-Releasing Hormone