This study is proposed to characterize nanocellulose extracted from sugarcane bagasse (SCB). For extracting cellulose, the bagasse was treated by chemical treatment. The nanocellulose was prepared using ball-milling with acid hydrolysis. Morphology of bagasse, cellulose and nanocellulose were characterized by scanning electron microscopy (SEM). The chemical composition of extracted cellulose and nanocellulose was investigated by Fourier-transformed infrared spectroscopy that verifying the removal of lignin and hemicellulose during cellulose isolation process from sugarcane bagasse. The crystalline structure of cellulose and nanocellulose were characterized by X-ray diffraction (XRD), indicating that the nanocellulose prepared by ball milling with acid hydrolysis has higher crystallinity than nanocellulose prepared only ball milling.