Detection of AMH protein in bovine serum and production of polyclonal antibodies for fertility diagnostic applications

This study aimed to isolate and characterize Anti-Müllerian Hormone (AMH) protein from Peranakan Ongole (PO) cows and to produce polyclonal antibodies for potential reproductive biomarker applications. Nine PO cows were selected based on ultrasonographic and rectal palpation examinations, which confirmed active follicular development. Serum samples were collected and analyzed for total protein concentration, ranging from 55.73 to 70.23 mg/mL. SDS-PAGE analysis revealed varied protein profiles, with 3 to 10 bands observed per sample, indicating physiological diversity among the subjects. AMH protein was isolated using electroelution following SDS-PAGE separation. Western blot results confirmed that the immunogenic AMH protein was detected at a molecular weight of 60–70 kDa, aligning with the expected size for bovine AMH. The purified protein was used for immunization in rabbits to produce polyclonal antibodies. The successful detection and specificity of the anti-AMH antibodies support their potential use in reproductive physiology studies and the development of immunodiagnostic tools.