UCSF

Differential scanning fluorimetry (DSF) is a technique used to determine a protein’s stability. In a typical DSF experiment, a purified protein is heated in the presence of a fluorescent dye and the resulting temperature vs fluorescence plot is used to calculate its apparent melting temperature. This technique can be used to study a wide range of phenomena, including binding of a protein to ligands or cofactors and the effects of point mutations or buffers on a protein’s relative stability. The advantage of DSF is that it can be used in low volume and high-throughput such that hundreds or thousands of conditions can be rapidly tested. This guide provides an introduction to DSF and guidance on assay development and data interpretation, including how to avoid common artifacts.