Objectives: This study aims to the development of nursing students’ attitudes scale toward pain assessment and establish its validity and reliability. Methods: This was a methodological study conducted in the 2017–2018 academic year. The sample consisted of 300 nursing students attending at the Department of Nursing of Gazi and Selçuk Universities. First, a 51-item draft was developed based on a literature review and interviews with 25 students. Five experts were consulted for content validity. The items were revised, and six items were removed based on their feedback. The 45-item final version was applied to participants. Afterward, exploratory and confirmatory factor analyses were performed to determine the scale structure. Test-retest reliability was determined on 190 participants selected randomly from the sample. Results: A 51-item draft was developed based on a literature review and interviews. Five experts were consulted for content validity, and six items were removed based on their feedback. The rotated principal component analysis revealed 15 items loaded on two factors. The total scale had internal consistency reliability (Cronbach’s alpha; α) of 0.918 and test-retest reliability of 0.738. Conclusion: Factor analyses showed that the scale had satisfactory construct validity and a two-factor structure. All α values were higher than 0.70, indicating that the scale had a satisfactory level of reliability. All in all, it is a valid and reliable scale that can be used to measure nursing students’ attitudes toward pain assessment.