Objective: Bicycling is an affordable way to increase access to employment, schooling, and services and an effective measure against obesity. Bikeshare programs can make bicycling accessible to diverse populations, but little evidence exists on their adoption in low-resource neighborhoods. Our study examined factors associated with bikeshare use in a metropolitan area in the southern United States. Methods: We performed a retrospective cross-sectional analysis of a database of clients (N=815) who rented a bicycle from Zyp Bikeshare in Birmingham, Alabama between October 2015 and November 2016. Individual-level variables included bike use frequency, average speed, total miles traveled, total minutes ridden, bike type (traditional vs electricity-assisted pedelec), membership type, sex, and age. Area-level data aggregated to Census tracts, proxies for neighborhoods, were obtained from the 2010 US Census after geocoding clients' billing addresses. Using exploratory factor analysis, a neighborhood socioeconomic disadvantage index (SDI) was constructed. Bikeshare station presence in a tract was included as a covariate. Multivariate linear regression models, adjusted for clustering on Census tracts, were estimated to determine predictors of bikeshare use. Results: In a multivariate regression model of individual and neighborhood characteristics adjusted for clustering, each decile increase in the SDI was associated with a 9% increase in bikeshare use (P<.001). Bikeshare use was also positively associated with speed (.1, P<.001), total miles (.008, P<.001), and pedelec use (1.02, P<.01). Conclusion: Higher neighborhood socioeconomic disadvantage is associated with higher bikeshare use. Bikeshare is a viable transportation option in low-resource neighborhoods and may be an effective tool to improve the connectivity, livability, and health of urban communities.