In biomedical research, rabbits are commonly sedated to facilitate a variety of procedures. Developing a sedation assessment scale enables standardization of levels of sedation and comparisons of sedation protocols, and may help in predicting sedation level requirements for different procedures. The goal of this study was to develop a rabbit sedation assessment scale using a psychometric approach. We hypothesized that the sedation scale would have construct validity, good internal consistency, and reliability. In a prospective, randomized, blinded study design, 15 (8 females, 7 males) healthy 1-y-old New Zealand white rabbits received 3 intramuscular treatments: midazolam (0.5 mg/kg; n = 6); midazolam (1.5 mg/kg)-ketamine (5 mg/kg; n = 7); and alfaxalone (4 mg/kg)-dexmedetomidine (0.1 mg/kg)-midazolam (0.2 mg/kg; n = 3). One rabbit received 2 treatments. A sedation scale was developed by using psychometric methods, with assessment performed by 6 independent raters who were blind to treatment. Final sedation scale items included posture, palpebral reflex, orbital tightening, lateral recumbency, loss of righting reflex, supraglottic airway device placement, toe pinch, and general appearance. The scale showed construct validity, good to very good interrater reliability for individual items (6 raters; intraclass correlation coefficient, 0.671 to 0.940), very good intrarater reliability (5 raters; intraclass correlation coefficient, 0.951 to 0.987), and excellent internal consistency (Cronbach α, 0.947). The sedation scale performed well under the conditions tested, suggesting that it can be applied in a wider range of settings (different populations, raters, sedation protocols).