A field study using 322 lactating dairy cows in seven commercial and two university herds was conducted to determine if treatment with estradiol benzoate 40 to 48 h after treatment with prostaglandin F2 alpha would enhance synchronization of estrus. Estrogen treatment tended to increase the proportion of cows in estrus within 5 d (synchronized) after prostaglandin treatment (66.9% versus 58.9%). Of synchronized cows, a greater proportion treated with estrogen (66.9%) were in estrus on d 3 than those not receiving estrogen (48.2%). First service conception rate (31.9%) and interval to second service (35.6 d) were not affected by treatment with estrogen. Milk progesterone was measured in university herds. More cows with milk progesterone concentrations greater than or equal to 8 ng/ml were synchronized (75.4%) than those with less than 8 ng/ml (63.3%). Treatment with estrogen increased synchrony of cows with high progesterone (90.3%) more than prostaglandin alone (60.0%). Based on progesterone concentrations at breeding and 22 to 24 d later, estimated conception rate was 58.7%, and net conception rate based on palpation was 41.3%. Tighter synchrony of estrus can be achieved by using estradiol benzoate 40 to 48 h after prostaglandin. Concentrations of milk progesterone might predict success of treatment.