Background: Nursing, as an essential component of contemporary healthcare systems, has undergone significant changes in recent decades, resulting in an increase in research across various fields, such as mental health, well-being, and the development of the discipline itself. Currently, gender stereotypes persist, impacting the social image of the profession and influencing the professional identity of nurses and consequently, their work performance and the quality of care provided. Both public and internal perceptions of the profession are critical factors in these dynamics. Objective: to adapt and validate the Nursing Attitude Questionnaire in a Spanish sample, as it has only been validated in two languages to date. Methods: This study presents descriptive and validity analyses of several measures utilized, such as Nursing Image (NIS), Medical Empathy (JSE-HP), Professional Values (NPVS) and Communication Skills (HP-CSS). Results: Confirmatory factor model analyses indicated that a 21-item hierarchical model provided a superior fit (χ2(df = 1.65, NFI = 0.954, SRMR = 0.065), suggesting the presence of a general factor accounting for five first-order factors. Internal consistency was deemed adequate for the total measures (α = 0.789), though variable for the subscales. Correlations among measures provided evidence of convergent and discriminant validity, highlighting consistent correlation patterns between Attitude toward Nursing and other variables. In addition, significant differences were observed in the Professionalism subscale according to sex, albeit with a medium-low effect size. Scales are presented to facilitate future interpretation of the results in similar samples. Conclusions: The scale validated in this study exhibited overall good psychometric performance, although certain limitations were identified within the subscales. The established scales represent a novel contribution, providing a valuable tool for the comparison of similar samples.