Research on cultural factors and substance use among Hispanic adolescents has focused primarily on acculturation, while specific core Hispanic values and attributes have received minimal attention. The objective of the current study was to examine the relationship between traditional Hispanic cultural assets and substance use among adolescents. A purposive sample of 225 Hispanic adolescents (47% male) aged 13 to 16 years were recruited from community venues (e.g., park, school, mall) in a predominately Hispanic neighborhood. Participants completed a survey to assess cultural factors (familism, simpatía, respeto, and ethnic pride) and substance use in the past 3 months (alcohol and drug). Point-biserial correlations revealed significant associations of alcohol and drug use with greater familism (family connectedness), simpatía (interpersonal relationship harmony), and respeto (respect). Two stepwise binary logistic regressions were performed to evaluate the independent association between the cultural factors and substance use. The interaction of gender with each cultural factor was examined in both analyses. Simpatía emerged as the only cultural factor independently associated with alcohol use. Greater simpatía was related to abstention from alcohol. Both simpatía and familism independently correlated with drug use. Stronger endorsements of simpatía and familism were associated with absence from drug use. Interactions between cultural factors and gender were not observed. Simpatía emerged as the strongest cultural asset that may confer protection against substance use. If replicated, our results suggest substance prevention programs targeting Hispanic adolescents may benefit from the inclusion of cultural assets in the intervention paradigm.