To deal with the turbulent environments, firms have endeavored to achieve greater supply chain collaboration. In researching the antecedents or the conditions that lead to or affect supply chain collaboration, prior studies focus on the use of interorganizational systems (IOS) but simplify or ignore its culture context. Although IOS use is necessary for supply chain collaboration to succeed, organizational culture must be taken into consideration simultaneously. Many supply chain collaborations fail due to incompatible corporate culture and the complexities involved. The objective of the study is to explore the impact of collaborative culture and IOS use on supply chain collaboration by examining a moderated mediation model. Data was collected through a Web survey of U.S. manufacturing firms. Structural equation modeling (LISREL) was used to analyze the data. Following the steps and procedures for implementing the latent variable interaction by orthogonalizing via residual-centered approach, the model was tested. The results indicate that collaborative culture enhances supply chain collaboration directly as well as indirectly by facilitating IOS use, which in turn improves supply chain collaboration. Thus, IOS use partially mediates the relationship between collaborative culture and supply chain collaboration. Surprisingly, the moderating effect of collaborative culture on the relationship between IOS appropriation and supply chain collaboration is not supported.