Objectives To evaluate the efficacy and safety of tacrolimus ointment 0.1% in adult and 0.03% in pediatric patients with moderate to severe atopic dermatitis in Malaysia. Methods This is an open-labeled and single arm multi-center study. 36 adult and 37 pediatric patients were enrolled. Tacrolimus ointment is applied twice daily for four weeks. The primary efficacy outcome is based on the Physician’s Global Evaluation of Clinical Response (PG) at Week 4. The secondary efficacy outcomes are Eczema Area and Severity Index (EASI) score, changes from baseline in individual scores of signs and symptoms and body surface area affected and Patients Assessment of Treatment Effects. Results Overall success rate were 97.1% and 91.2% in the adult and pediatric groups respectively. The decline in EASI, percentage of total BSA affected and patient’s assessment of pruritus were significant (P<0.001). Of adults and pediatric patients, 97.2% and 75.7% respectively reported adverse effect. The most common adverse effect reported was skin burning sensation in 91.7% adult patients and pruritus in 67.6% pediatric patients. Conclusion Tacrolimus ointment 0.1% in adult and 0.03% in pediatric patients is effective for the treatment of moderate to severe atopic dermatitis in Malaysia.