10.4225/03/5a1e35aab0544 David Lindsay Healy David Lindsay Healy Human prolactin physiology Monash University 2017 prolactin 2017-11-29 04:20:56 Thesis https://bridges.monash.edu/articles/thesis/Human_prolactin_physiology/5643313 The overall aim of the work to be presented in this thesis was to investigate the biology of the protein hormone prolactin (PFL) in two areas of reproductive endocrinology: hypothalamic-pituitary control of PRL secretion and the possible origin of amniotic fluid PRL. For these studies, a reliable homologous PRL double antibody radioimmunoassay was developed using materials supplied by the NIAMDD. Serum PRL concentrations in 60 normal men were 12.8 ± 3.2 ng/ml (mean± 2 SO) and in 24 premenopausal women 13.6 ± 5.4 ng/ml. Depending onthe iodination procedure used, the mean assay sensitivity was 0.55 -0.61 ng/ml, the within-assay precision (expressed as a coefficient of variation) was between 5.6 - 6.6 per cent and the between-assay precision for medium pool samples was 16 - 19%. Factors crucial to the development of a reliable assay were the use of minimal oxidation conditions for PRL iodination, complete separation of the iodinated species so formed, use of a highly dilute PRL antiserum to increase assay sensitivity, careful titration of the amount of non-immune serum used to increase ant genic bulk for the second antibody step and a long incubation time to ensure maximal immunoprecipitation. [...]