This study presents serum concentrations of pregnancy-associated glycoprotein (PAG), oestrone sulphate (E1-S) and progesterone (P4), and the effects of some dam and foetus-related factors on these profiles during gestation in Borana and crossbred cattle. The PAG concentrations at 4th week post-conception ranged from 1.5-5.5 and 2.1-4.7 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The mean PAG concentrations increased progressively from 4th to 33rd week of gestation (from 3.3-173 ng/ml for Borana and 4.2-240 ng/ml for crossbred cattle) and reached peak around calving. Breed, parity status, dam body weight, foetal sex and foetal birth weight significantly influenced the PAG concentrations. After delivery, the PAG concentrations declined steadily to 5.7 ng/ml in Borana (n = 7) and 3.9 ng/ml in crossbred (n = 6) cattle 10 weeks post-partum. The serum E1-S concentrations at 17th week of pregnancy ranged from 0.3-2.6 and 0.9-5.7 ng/ml in Borana (n = 8) and crossbred (n = 9) cattle, respectively. The mean E1-S concentrations increased progressively from 17th to 33rd week of gestation (from 1.1-4.6 ng/ml for Borana and 2.7-10.8 ng/ml for crossbred). Breed, parity status, dam body weight and foetal sex significantly influenced E1-S concentrations. The P4 concentrations at 4th week of pregnancy ranged from 3.2-5.1 and 1.7-8.9 ng/ml in Borana (n = 6) and crossbred (n = 8) cattle, respectively. The P4 level remained elevated throughout pregnancy. This study indicated that the serum PAG and P4 concentrations at 4th and E1-S approximately 17th week of pregnancy were above the cut-off value for pregnancy test and the hormonal profiles observed were comparable to the previous reports. Furthermore, the PAG and E1-S profiles were considerably influenced by factors such as breed, weight and parity status of the dam, and foetal sex and foetal birth weight (only PAG).