Measuring the KD of Protein–Ligand Interactions Using Microscale Thermophoresis

Shih Chia Tso, Chad A. Brautigam

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Microscale thermophoresis (MST) has become a widely used technique to determine the KD or EC50 of protein–ligand interactions. The method exploits the tendency of macromolecules to migrate along a thermal gradient (i.e., thermophoresis). Differences in thermophoresis as a function of the liganded state of a macromolecule can be measured and assembled into a binding curve that can be analyzed to yield KD. In this protocol, we outline a simple experiment designed for new MST users, with the goal of using readily available, inexpensive materials to plan, execute, and analyze an MST experiment.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages161-181
Number of pages21
DOIs
StatePublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2263
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Affinity measurement
  • K
  • Microscale thermophoresis
  • Protein–ligand interactions
  • Protein–protein interactions

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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