In this study we evaluated the psychometric properties of the Homophobic Bullying Scale in a sample of Spanish adolescents. The study comprised 791 adolescents between 12 and 17 years old (M= 13.96, SD= 1.18) who completed this questionnaire along with the Modern Homophobia Scale that measures homophobic attitudes. The results of the factor analyses confirmed a structure of four intercorrelated factors: 1) observation of homophobic bullying behaviors against gay boys, 2) observation of homophobic bullying behaviors against lesbians, 3) perpetration of homophobic bullying against gay boys, and 4) perpetration of homophobic bullying against lesbians. The questionnaire showed good reliability and perpetration scores were associated with homophobic attitudes. Higher prevalence of bullying was found against gay boys than against lesbian girls. These results show that this is a simple and valid questionnaire for the evaluation of homophobic bullying among Spanish adolescents.