Indirect Detection of Ligand Binding by Thermal Melt Analysis.

A thermal shift assay (TSA) involves measuring the effect of a compound on the thermal stability of a protein as an indirect measure of ligand binding. In this chapter, we provide a protocol for a conventional TSA with recombinant/purified proteins using differential scanning fluorimetry (DSF), followed by a protocol for a Cellular Thermal Shift Assay (CETSA), which measures the soluble cellular protein remaining after a transient heat shock of live cells to detect intracellular ligand binding.