The effect of flow rate and perfused load on insulin absorption was studied in isolated proximal rabbit tubules. In one series of experiments tubules were perfused for 30 min with 125I-[A14]insulin at 3.6 ng/ml and at flow rates varying between 2.2 and 43.3 nl/min, thus varying both flow rate and the perfused load. In a second series of experiments the tubules were exposed to increasing loads of insulin at a fixed flow rate of 15 nl/min. In a third series of experiments two groups of tubules were exposed to identical perfused loads at two different flow rates. The results showed that absorption is linearly correlated with the perfused load in the ranges of flow rates and concentrations examined. The perfused load determined the magnitude by which the insulin was reabsorbed, whereas the flow rate determined the efficiency (fractional absorption) by which the protein was reabsorbed. At flow rates less than 5 nl/min the fractional absorption was greater than 65%, and at flow rates greater than 30 nl/min fractional absorption was less than 40%. Thus, there is an inverse relationship between flow rate and fractional absorption. The described effects of changes in flow rate and perfused load are suggested to be caused by concomitant changes in the mean luminal concentration.