From the experimental point of view, the most direct way to validate a docking pose prediction is by determining the experimental protein structure in complex with the ligand, typically through X-ray crystallography or NMR spectroscopy. Also, assays such as surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), and enzyme inhibition studies can provide experimental evidence for the binding affinity and kinetics of the ligand-protein interaction, which can be compared to docking predictions. Another experimental technique is site-directed mutagenesis that can be used to alter specific amino acids within the binding site. The effects of these mutations on ligand binding can validate the importance of predicted interactions .