This article aims to construct and validate an instrument for measuring the factors of organizational culture that influence cooperation in technological innovation between research institutes and firms, nominated “Evaluation of Organizational Culture for Cooperation in Technological Innovation (EOC-CTI)”, applied to the reality of Brazilian organizations. The sample was composed of 324 respondents from 58 large and medium firms and 41 public and private research institutes, from various locations in Brazil. The method adopted was the Exploratory Factor Analysis. Among the main contributions, an instrument that expands the discussions about the values and management practices for cooperation in technological innovation based on organizational culture is proposed. For firms and research institutes, there is the proposition of values and systematic management practices that contribute to the achievement of results through cooperation in technological innovation.