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Introduction: End-stage renal disease (ESRD) is an advanced stage of chronic kidney disease requiring hemodialysis (HD). The long-term efficacy of HD in ESRD patients highly depends on treatment adherence. Objectives: This study aimed to validate the health action process approach (HAPA) questionnaire to predict treatment adherence in HD patients. Patients and Methods: This cross-sectional study was conducted in teaching hospitals in Shiraz during 2018. A total of 220 patients with ESRD under HD were selected using convenience sampling method. Furthermore, the validity, clarity, and comprehensiveness of the questionnaire were validated by a group of patients and experts. Then the exploratory factor analysis (EFA) was performed, and the reliability was determined using Cronbach’s alpha. Internal consistency was assessed using test-retest method (one-month interval) and calculating intra-class correlation coefficient (ICC) index. Results: Content validity index (CVI) and content validity ratio (CVR) were obtained 0.98 and 0.95 respectively indicating adequate content validity. Six constructs (risk perception, action self-efficacy, behavioral intention, planning, maintenance self-efficacy and recovery self-efficacy) were extracted using EFA. These constructs explained 51.4% of total variance. The Cronbach’s alpha of different constructs ranged from 0.68 to 0.82. Furthermore, the ICC ranged from 0.67 to 0.78 indicating an acceptable internal consistency. Conclusion: The HAPA questionnaire is a valid and reliable tool for assessing treatment adherence in HD patients. Further studies are recommended on larger sample sizes and other Iranian populations.