Recently, the technology of triboelectric nanogenerator (TENG) has been developed with expecting to compensate the batteries by converting mechanical energy into electricity. It also has brought numerous changes in the medical uses, especially in implantable/wearable technologies. The present study aims at the preparation of natural silk in form of silk fibroin (SF) film for possible applications as transparent TENG. Silk cocoon can be transformed into SF solution by Ajisawa’s method. Then, SF film was further fabricated by a simple casting process. The obtained SF film was analyzed by using Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy (ATR-FTIR), Scanning Electron Microscopy (SEM) and UV-visible Spectrophotometer (UV-Vis). ATR-FTIR results demonstrated the N-H bonding, C=O bonding, and C-N bonding, confirming the Amino group in SF molecules. The SEM image shows the smooth surface morphology and the UV-Vis result shows highly transparent in the visible light. By fabricating devices in vertical contact separation mode, the present SF-TENG provides an output voltage and current of ~67.8 V and ~4.36 µA. The maximum power of ~123.4 µW was obtained at the load resistance of 7 MΩ. The electrical output performance generated from this SF-TENG has excellent performance for utilizing as a power source of small electronic devices.