Teacher collaboration is considered an important determinant when implementing differentiated instructional practices. However, most research has extensively explored such constructs using global measures, while overlooking the fact that teacher collaboration is a multidimensional construct. Against this background, this study aimed to explore the different teacher collaboration categories (exchange, synchronization and co-construction) and their impact on teachers’ differentiated instructional practices. In addition, this study took into consideration important variables for teachers’ differentiated instructional implementation, such as self-efficacy, attitudes, and the contextual factor of school track. To respond the research questions, descriptive statistics, one-sample t-tests, mixed analysis of variance with and hierarchical linear regressions were conducted. Results from a mixed analysis of variance indicated that teachers mainly collaborate by means of less demanding and less intensive cooperative practices, such as the exchange of teaching materials and content-related information. Additionally, findings from a hierarchical linear regression indicate that more demanding practices of collaboration, such as synchronization and co-construction, are significantly associated with teachers’ differentiated instructional practices. Implications of the results as well as further lines of research are discussed.