Peripheral plasma samples were obtained during pro-oestrus and oestrus from two adult Beagle bitches and from one animal (bitch 124) during its first oestrous cycle. The animals were housed in a constant environment kennel and each showed a normal oestrous cycle as judged by clinical appearance, vaginal cytology and behavioural tests; in addition, a rise in plasma progesterone was noted during late oestrus and early metoestrus (Christie, Bell, Horth & Palmer 1971). Plasma oestrogens were measured by the radioimmunoassay technique of Abraham (1969), with modifications. No separation of oestradiol and oestrone was undertaken and the results are thus an estimate of total immunoreactive oestrogens. The water blank for the method was 16 ± 6·5 (S.D.) pg/ml while the solvent blank was 19 ± 4·7 pg/ml. The results were corrected for the solvent blank. The recovery of oestradiol added to water was 102 ± 12 (S.D.)%, and to plasma from a bilaterally ovariectomized, hemi-adrenalectomized.