We aimed to determine the effect of timed artificial insemination (TAI) protocols on the pregnancy rate per insemination and pregnancy loss compared with AI performed at detected estrus in dairy cows and Korean Hanwoo cattle under heat stress. In dairy cattle, 1,250 sets of data that underwent AI during heat stress (temperature-humidity index ≥ 72) were categorized according to their TAI protocols or as controls: 1) PGF2α-36 h-estradiol benzoate (EB)-36 h-TAI (PG-EB group, n = 113); 2) GnRH-7 days-PGF2α-56 h-GnRH-16 h-TAI (Ovsynch group, n = 455); or 3) GnRH-6 days-Ovsynch (G6G group, n = 136). The remaining cows underwent AI at detected estrus (AIDE group, n = 546). The probability of pregnancy per AI 45 days after AI was higher (P < 0.01) in the PG-EB (odds ratio [OR]: 1.68), Ovsynch (OR: 1.48), and G6G (OR: 1.79) groups than in the AIDE group. However, the prevalence of pregnancy loss between 30 and 45 days after AI did not differ among the groups. In Hanwoo cattle, 617 sets of data inseminated artificially under heat stress were categorized into AIDE (n = 281), PG-EB (n = 194), and combined Ovsynch or G6G (n = 142) groups. The probability of pregnancy per AI 45 days after AI and the prevalence of pregnancy loss between 30 and 45 days after AI did not differ among the groups. Thus, implementation of a TAI protocol (PG-EB, Ovsynch, or G6G) in dairy cows under heat stress improves the pregnancy rate per AI versus AIDE, whereas there is no beneficial effect of TAI on the pregnancy rate of Hanwoo cattle under heat stress.