The detection of Highly Pathogenic Avian Influenza (HPAI) A(H5N1) in dairy cattle in the United States has raised concerns about human exposure. This study evaluated the efficacy of various doses of an inactivated H5 AI vaccine in cattle and assessed antibody transfer in milk against a recent bovine isolate of HPAI A(H5N1, clade 2.3.4.4b). Calves were inoculated with different vaccine doses, while lactating cows received the vaccine four weeks later. The humoral immune response was measured using the Hemagglutination Inhibition (HI) test and ELISA. Results showed a dose-dependent immune response, with higher doses producing stronger and more sustained antibody levels. Group 1 maintained a stable HI titer of 6 log2, while Groups 2, 3, and 4 peaked at 8, 9, and 9 log2, respectively, by the fourth week post-vaccination. Milk antibody transfer was observed, with strong positive responses in milk samples by the second week post-vaccination. The ID Screen ELISA demonstrated higher sensitivity for detecting antibodies in milk compared to serum. The immune response to the AI vaccine differed from responses to other vaccines used in cattle such as Foot and Mouth Disease Virus (FMDV) and Lumpy Skin Disease Virus (LSDV), indicating the need for optimizing vaccine dosage and formulation, including adjuvant and antigen content. Future research should extend the monitoring period, increase sample sizes, and explore different vaccine formulations to develop effective vaccination strategies for cattle. These findings highlight the potential for using inactivated H5 AI vaccines in cattle to enhance immune protection and facilitate antibody transfer through milk.