Ag grain boundary (GB) diffusion in Cu and Cu-0.2 at.% Ag alloy was measured in the B and C regimes and the equilibrium segregation factor s was determined for the pure metal and the alloy. In the C regime, Ag GB diffusivity Ag gb D in the Cu-0.2 at.% Ag alloy was directly measured and it was shown that Ag gb D is similar to the Ag GB diffusivity in pure Cu. The B regime penetration profiles are curved in both pure Cu and the Cu-Ag alloy. In order to elucidate the reasons of this curvature, Ag GB diffusion was measured in pure and alloyed Cu bicrystals. It was shown that in the present experimental conditions non-linear segregation affects the Ag penetration profiles in bicrystals, whereas linear segregation occurs in pure Cu polycrystals due to the different levels of Ag concentration present in the GBs. The effect of Ag alloying of Cu poly-and bicrystals depends remarkably on the temperature and produces a systematic variation of the profile shape and of the measured diffusion coefficients. As a result, the segregation isotherm of Ag in Cu GBs was determined.