Previous empirical research on tolerance suffers from a number of shortcomings, the most serious being the conceptual and operational conflation of (in)tolerance and prejudice. We design research to remedy this. First, we contribute to the literature by advancing research that distinguishes analytically between the two phenomena. We conceptualize tolerance as a value orientation towards difference. This definition—which is abstract and does not capture attitudes towards specific out-groups, ideas, or behaviors—allows for the analysis of tolerance within and between societies. Second, we improve the measurement of tolerance by developing survey items that are consistent with this conceptualization. We administer two surveys, one national (Sweden) and one cross-national (Australia, Denmark, Great Britain, Sweden, and the United States). Results from structural equation models show that tolerance is best understood as a three-dimensional concept, which includes acceptance of, respect for, and appreciation of difference. Analyses show that measures of tolerance have metric invariance across countries, and additional tests demonstrate convergent and discriminant validity. We also assess tolerance’s relationship to prejudice and find that only an appreciation of difference has the potential to reduce prejudice. We conclude that it is not only possible to measure tolerance in a way that is distinct from prejudice but also necessary if we are to understand the causes and consequences of tolerance.