The experiment examined patterns of mammary uptake of individual AA when graded amounts of calcium caseinate (0, 177, 362, and 762 g/d) were infused duodenally into four lactating cows. Six blood samples were collected over 12 h from the subcutaneous abdominal vein and the carotid. Mammary blood flow was measured by an ultrasonic flow probe implanted around the external pudic artery. Infusions of casein linearly increased the arterial concentrations of all essential AA and several nonessential AA (Pro, Tyr, Orn, and Cit) and increased, or tended to increase, linearly the mammary arteriovenous differences of all AA except Glu and Ala. Absorption ability of the mammary gland was not reduced in vivo. Relationships between mammary arteriovenous differences and arterial concentrations were positive and linear in every cow for all AA except Asn, Ser, Gly, and Ala. Some essential AA (Lys, Arg, and branched-chain AA) were therefore taken up in excess of their output into milk proteins, but others (His, Thr, Met, and Phe) were almost exclusively extracted by the udder in a direct ratio to their output. As infusions of casein increase, Phe becomes probably the most critical AA for milk synthesis.