Fig. 2
From: Target identification of small molecules: an overview of the current applications in drug discovery
The standard experimental protocol for the photoaffinity method. 1. Synthesizing the photoaffinity probe. 2. Using photoaffinity probes on cells or cell lysates and allowing them to bind to their target. 3. Ultraviolet (UV) light is subsequently employed to activate the covalent crosslinking of the probes with the target proteins in the treated samples (cells or cell lysates). (If the experiment is performed on live cells, the cells are lysed after the UV exposure) proteins. 4.-Using streptavidin, the complex (target + probe) would be extracted. 5. Removing the unbound proteins by washing. 6. The target proteins can be analyzed using SDS-PAGE and identified through protein digestion and mass spectrometry