The effects of suckling on secretion of luteinizing hormone, cortisol and transcortin were investigated in anovulatory postpartum cows. On d 35 postpartum, calves were separated from 12 cows to prevent suckling and eight calves continued to suckle their dams ad libitum. Between 35 and 41 d postpartum, samples of jugular blood were collected every 15 min for two periods of 6 h/d. In non-suckled cows, frequency of pulses and basal luteinizing hormone increased but amplitude of pulses did not change. Concentrations of total cortisol in serum of cows were not altered during 3 d after weaning calves and did not differ among intervals before, during and after a suckling event. Affinity of transcortin for cortisol was not affected by postpartum interval or treatment. Capacity of transcortin to bind cortisol tended to increase after weaning. We found no evidence to support the hypothesis that suckling reduces binding capacity of transcortin or increases unbound cortisol. Differences in preovulatory secretion of luteinizing hormone between suckled and non-suckled cows could not be accounted for by differences in secretion of cortisol. In beef cows that are fed to satisfy requirements for energy and have average body condition, we conclude that negative modulation of luteinizing hormone by suckling is not mediated by cortisol.