Concealed antigens from the adult of Hyalomma anatolicum anatolicum were isolated by immunoaffinity chromatography using anti-gut IgG as ligands. The bound antigens were characterized by SDS-PAGE and 2 proteins of 106.7 and 68 kDa were identified. Immunization of crossbred calves (Bos taurus x Bos indicus) with 1.7 mg of the bound antigens adjuvanted with FCA/IFA animal decreased 74. 4 and 52.2% population of nymphs and adults respectively. Besides, a significant percentages of abnormally fed larvae (P < 0.001) and nymphs (P < 0.05) was dropped from the immunized animals. Western blot analysis detected protein band of 68 kDa as the antigen responsible for the induction of resistance in the host.