The military is a work environment in which the underrepresentation of women and the presence of gender prejudices continue to exist. The present study validated the Ambivalent Sexism Inventory (ASI) for the Croatian military population. To determine the ASI scale’s basic metric characteristics, data were collected from a sample of 895 active-duty military personnel (445 men and 450 women). The study results determined satisfactory measurement characteristics for the ASI scale and confirmed the ambivalent sexism construct with its basic dimensions. Sexism in the military environment was found to a moderately high degree, and gender differences were observed. Three types of sexism endorsement were identified in both subsamples (egalitarian, moderate egalitarian and traditional for women, and moderate egalitarian, traditional and hostile for men), with additional differences detected in their socio–demographic and professional characteristics. The findings support the apparent exposure of women to sexism in the military environment, and suggest the need to raise awareness of the negative impact of gender prejudice on gender relations in the military.