HIV stigmatizing attitudes are embedded in social context, making it important to develop culturally specific tools for accurate measurement. The goal of this study was to develop and evaluate the psychometric properties of the HIV Stigmatizing Attitudes Scale (HSAS) in Moshi, Tanzania. Items were adapted based on a scale developed by Visser et al. which was one of the first to measure HIV stigmatizing attitudes in the general population (i.e., people not living with HIV). Items were translated into Swahili and modified with iterative feedback. The HSAS was administered to participants (N = 1494) in an HIV stigma reduction intervention study at two antenatal care clinics in Moshi, Tanzania. The HSAS was found to have strong domain coherence and high reliability based on Cronbach’s alpha, Omega 6 coefficient values, and the composite reliability coefficient, and high validity based on content-oriented evidence, relations to other variables, and response process. Factor analysis revealed a two-factor structure (Moral Judgment and Interpersonal Distancing), consistent with the original Visser scale. The HSAS provides a robust way to measure HIV stigma in the Tanzanian context and can be culturally adapted to other settings.