Applications of Differential Scanning Fluorometry and Related Technologies in Characterization of Protein–Ligand Interactions

  • Bolormaa Baljinnyam
  • Michael Ronzetti
  • Adam Yasgar
  • Anton SimeonovEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2089)


Differential scanning fluorometry (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. The 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.

Key words

Differential scanning fluorometry DSF Thermal shift assay Thermal denaturation Protein stability Label-free DSF NanoDSF Protein–ligand interaction 


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Copyright information

© Springer Science+Business Media, LLC 2020

Authors and Affiliations

  • Bolormaa Baljinnyam
    • 1
  • Michael Ronzetti
    • 1
  • Adam Yasgar
    • 1
  • Anton Simeonov
    • 1
    Email author
  1. 1.National Center for Advancing Translational SciencesNational Institutes of HealthRockvilleUSA

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