The chopper chelator cuprizone serves as a valuable chemical tool to induce consistent and reproducible demyelination in the central nervous system. However, the daily preparation of fresh cuprizone powder mixed in finely ground rodent chow might well be a particular health problem. Alternative methods, such as the fabrication of cuprizone-containing pellets, are available. The effectiveness of this method is, however, not known. In the present study, we compared whether intoxication of C57BL/6 mice with 0.25% cuprizone mixed into ground rodent chow does induce demyelination to a similar extent compared to a cuprizone-pellet intoxication protocol. We found that feeding of 0.25% cuprizone in ground chow provides a strong, well-defined, and reproducible demyelination along with increased accumulation of microglia and axonal damage in the corpus callosum, whereas all analyzed parameters were significantly less distinct in mice fed with cuprizone-containing pellets at an equivalent concentration of cuprizone at week 5. Even a higher concentration of cuprizone in pellet formulation was less potent compared to do so. This study illustrates that the established protocol of cuprizone intoxication (i.e., mixed in ground rodent chow) is the gold standard method to achieve consistent and reproducible demyelination. Why cuprizone loses its effectiveness in pellet formulation needs to be addressed in subsequent studies.