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Following the observation that the concentration of progesterone in goats' milk differs appreciably according to the specificity of the antiserum used in a non-extraction (direct) radioimmunoassay, experiments were carried out to find an explanation for these results. Milk and plasma samples were collected during the oestrous cycle and during an equivalent period of pregnancy after a fertile mating. Samples were analysed by a direct radioimmunoassay using two antisera, 18/3 which is highly specific for progesterone and 465/6 which is less specific, and by radioimmunoassay of fractions isolated by thin-layer chromatography (TLC). Values obtained for milk and plasma samples collected during the oestrous cycle and early pregnancy were similar, except that values for milk samples measured with antiserum 465/6 were higher in pregnancy compared to those obtained during the oestrous cycle. Values obtained for milk and plasma with antiserum 465/6 were significantly higher than those obtained with 18/3 (P<0·001). After TLC this difference was found to be due principally to the presence of compound(s) with chromatographic properties identical to 5-pregnanedione(s). A comparison of the concentration measured in arterial and mammary venous plasma and in milk showed that about 25% of progesterone (5·7 nmol/min) was extracted by the mammary gland, and that substantial amounts of immunoreactive metabolites of progesterone are secreted into milk with only small quantities being transferred into mammary vein plasma.