The purpose of this study was to create a Mid-career Generalist Nurses Leadership Scale (MGNLS) and examine its reliability and validity. We created 54 MGNLS items based on interview data and a literature review. We asked 61 head nurses and 249 nurses in their first to fourth years of work at 15 hospitals to complete a questionnaire survey (N = 310). We then conducted exploratory factor analysis. The factor analysis revealed 33 MGNLS items and 6 factors (assertive transmission ability to examine problems, junior-centered growth support behavior, practice of workplace learning, challenge to change, collaboration and self-control ability, and reflection behavior as a leader). The reliability of the measure was ensured by confirming the internal consistency, and the concept of the structure was supported. The validity of the construct concept of the MGNLS was confirmed by examining the relationship between servant leadership and team leadership and by examining the impact of MGNLS score on a team member’s job satisfaction and teamwork. Thus, the data supported the reliability and validity of MGNLS.