The proportions of milk stored within the gland cistern (cisternal milk) and within secretory alveolar tissue (alveolar milk) were determined at various intervals after milking in peak lactation cows and late lactation cows. The rate of milk secretion remained constant up to 12 h in both groups. Cisternal milk remained low (600 g or less) until after 4 h in both groups, then increased gradually to values at 12 h of 5.08 (peak lactation) and 2.60 kg (late lactation). Alveolar milk exceeded cisternal milk in both groups and at all time points up to 12 h, and was greatest in the peak lactation cows. A further measurement was made at 20 h in the late lactation group. Milk secretion rate remained constant between 12 and 20 h and most of this additional milk accumulated in the cistern. Alveolar fraction, defined as alveolar milk as a proportion of total milk, ranged from 0.43 to 0.94 at 8 h. There was a highly significant correlation between 8 h and 20 h alveolar milk fraction results, but not between 1 h and 8 h values, nor between alveolar milk at 0 h (effectively residual milk) and alveolar fraction at any time point. The milk volume results were used to construct a mathematical model describing milk flow between cisternal and alveolar compartments, and hence the pattern of milk accumulation. The results are discussed in relation to current hypotheses regarding local autocrine control of milk secretion by a feedback inhibitor in milk.