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Penelitian ini bertujuan mengevaluasi tingkat keberhasilan IB semen beku sexing Y. Penelitian menggunakan 39 sapi Peranakan Ongole dan 77 sapi Persilangan Limousin yang di IB secara double dosis. IB menggunakan semen beku spermatozoa Y dengan metode pemisahan Sentrifugasi Gradien Densitas Percoll (SGDP) sapi Limousin yang diproduksi oleh Balai Besar Inseminasi Buatan (BBIB) Singosari. Teknik deposisi semen adalah 4+ yaitu pada posisi cornua uteri selanjutnya dilakukan IB pada jam ke 2 dan ke 8. Untuk akseptor dilakukan penyuntikan BioATP+ dan pemberian pakan konsentrat 3 kg per hari selama 3 hari setelah IB. Variabel penelitian meliputi Non Return Rate (NRR), Service Per Conception (S/C), dan Conception Rate (CR). Hasil penelitian menunjukkan bahwa IB sapi Peranakan Ongole dan sapi Persilangan Limousin yaitu NRR1 sebesar 82,05% dan 89,61%, nilai NRR2 sebesar 76,92% dan 84,42%, nilai CR sebesar 58,97% dan 74,03%, serta nilai S/C sebesar 1,78% dan 1,46%. Kesimpulan penelitian adalah IB menggunakan semen beku hasil sexing Y sapi Limousin pada sapi persilangan Limousin memiliki tingkat keberhasilan yang lebih tinggi dibandingkan sapi Peranakan Ongole. (The success rate of artificial insemination using frozen sexing semen of different breed cow) ABSTRACT. This study aimed to evaluate the success rate of AI frozen semen sexing Y. The material in this study used 39 Filial Ongole and 77 Limousin crossbred cattle in AI in double dose. The spermatozoa used were Y frozen semen of Limousin bull with the Percoll Gradient Density Centrifugation (SGDP) separation method produced by the Singosari Center for Artificial Insemination. The semen deposition technique was 4+, which has the position of the cornua uteri (deep Insemination), then AI implemented with a double dose at the 2 nd and 8 th hours after estrous. The cow acceptor was injected with BioATP+ and feed by 3 kg concentrate per day for three days after AI. Research variables include Non-Return Rate (NRR), Service Per Conception (S/C), and Conception Rate (CR). The results showed that AI of Peranakan Ongol and Limousin crossbred cattle on NRR1, NRR2, CR values, and S/C values respectively was 82.05% and 89.61%; 76.92% and 84.42%; 58.97% and 74.03%; 1.78% and 1.46%. In conclusion, AI using frozen semen from sexing Y Limousin cattle in Limousin crossbred cows had a higher success rate than Filial Ongole cattle.