Mad1 gene was ubiquitously exist in Metarhizium anisopliae, and responsible for adhering to insectsurface. To obtain high purity protein of MAD1 and its polyclonal antibody for studying the function of Mad1 gene, itwas cloned from M. anisopliae by RT-PCR, and then connected into the prokaryotic expression vector for the expres-sion. The protein was induced by IPTG and purified by metal chelate affinity, and then rabbits were immunized forantibody preparation. The specificity of polyclonal antibody were detected by Western-blot and indirect ELISA. Theresults indicated that high specificity antibody was obtained. The antibody titer was 1:12800, which will give goodexperimental materials for Mad1 gene function research and detection.