MST (Microscale Thermophoresis) Analysis

MST (Microscale Thermophoresis) is an optical method for characterizing biomolecules, involving the directional movement of particles within a microscopic temperature gradient. It utilizes infrared lasers to establish precise temperature gradients at the microscale, and studies molecular interactions by monitoring the thermophoretic movement (directional migration from high-temperature to low-temperature regions) of fluorescently labeled molecules within this gradient field. When a ligand binds to a target molecule, changes occur in the complex's hydration layer, charge, and molecular weight, leading to variations in the thermophoretic rate. By recording changes in fluorescence signals at different ligand concentrations, the binding affinity (Kd) can be quantitatively determined. This method allows for direct detection in solution, eliminating the need for sample fixation, and offers advantages such as low sample consumption and wide applicability.