Gelatin is one of the most popular biopolymers widely used in food industries. It has been mainly derived from porcine skins and bovine bones. However, gelatin from fish has drawn extensive interest due to the requirements for Halal and Kosher ingredients for food product development and rising concern of consumers about BSE in commercial mammalian gelatin products. Therefore, the extraction of gelatin from pangasius (Pangasianodon hypophthalmus) bone was investigated. The study aims to determine the optimal conditions for producing a good quality of gelatin and efficient utilization of pangasius by-products as well with four experiments (i) the effects of concentration of NaOH (0.2, 0.4 and 0.6%) and treatment time (22, 24 and 26 hours); (ii) the effects of HCl concentration (2, 4 and 6%) and treatment time (12, 24 and 36 hours); (iii) the effects of extraction time (6, 9, 12 and 15 hours) at 70-80oC and (iv) the effects of drying time (24, 48, 72 and 96 hours) at 55-60oC on quality of gelatin. The extraction conditions and their physic-chemical properties were compared to commercial gelatin from porcine skin. The results showed that protein and ash contents of gelatin from pangasius bone were 25.3% and 3.0%, respectively as 0.4% NaOH and treatment time for 26 hours, 4% HCl and treatment time for 24 hours were applied. The best viscosity was 5.18 mPa.s as applying an extraction time for 12 hoursat 70-80oC. Moisture and yield of extracted gelatin were 10.4% and 3.58%, respectively as gelatin solution dried for 48 hours at 55oC. Extracted gelatin from pangasius bone showed lower protein, ash and moisture contents, L* value while higher viscosity and gel strength compared to commercial gelatin.