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Updated: Aug 8, 2026

Determination of Protein-ligand Interactions Using Differential Scanning Fluorimetry
Published on: September 13, 2014
Applications of Differential Scanning Fluorimetry and Related Technologies in Characterization of Protein-Ligand
Bolormaa Baljinnyam, Michael Ronzetti, Adam Yasgar
1National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA. asimeono@mail.nih.gov.
Abstract:
Differential scanning fluorimetry (DSF) is an efficient and high-throughput method to analyze protein stability, as well as detect ligand interactions through perturbations of the protein's melting temperature. This method monitors protein unfolding by observing the fluorescence changes of a sample, whether through an environmentally sensitive fluorophore or by intrinsic protein fluorescence, while a temperature gradient is applied. Here, we describe in detail how to develop and optimize DSF assays to identify protein-ligand interactions, while exploring different buffer and additive conditions. Analysis of the data and further applications of the method are also discussed.
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