Microscale thermophoresis (MST) is a technique for biophysical analysis of biomolecular interactions. MST is based on the detection of temperature induced target fluorescence changes as a function of the concentration of non-fluorescent ligands. MST can be used with almost all kinds of target molecules, if they are fluorescent or can be labeled with fluorescent dyes. The diversity of ligands is greater. The fluorescent target molecule keeps a constant concentration, and the non-fluorescent ligand molecule titrates in a large concentration range.
MST is carried out in a thin capillary in free solution, thus providing near natural conditions (no immobilization in any buffer, even in complex biological liquids) and maintenance free instruments. In MST experiment, infrared laser can cause micro temperature gradient, and detect tric and thermophoresis. Tric depends on the microenvironment of the fluorophore and usually changes during binding events. The movement of molecules in a temperature gradient, thermophoresis, depends on three parameters that usually vary with the interaction. Therefore, the dose-response curve can be obtained by plotting the whole MST signal relative to the ligand concentration, from which the binding affinity can be deduced.
Figure 1. The principle of MST technique. (Wikipedia)
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