Spatial scale plays a crucial role in the assessment and management of ecosystem services (ES), yet explicit information for identifying and understanding the scale effect on ES supply remains limited. In an attempt to detect scale effect on ES supply from a comprehensive perspective, this study developed a framework for integrating scale effect in three aspects, including individual ES patterns, pairwise ES interactions, and ecosystem service bundles (ESB). The framework was tested in Xilinhot, a prairie landscape city of Inner Mongolia, at four different levels of spatial scale. The results indicated that, most ES showed a decreasing clustering at coarser scales in terms of spatial pattern. At the same time, coarser scales resulted in fewer trade-offs and stronger synergies between pairwise ES. The identification of ESB varied greatly with scale, and this change reflected in the composition of ES variables and spatial distribution of bundles. We attributed the scale effect of the above three aspects to differences in social-ecological factors and their driving mechanisms at different scales. This comprehensive framework could support local managers to coordinate the management of multiple ES at different scales.