Microscale Thermophoresis as a Sensitive Method to Quantify Protein: Nucleic Acid Interactions in Solution

Part of the Methods in Molecular Biology book series (MIMB, volume 815)


Microscale thermophoresis (MST) is a new method that enables the quantitative analysis of molecular interactions in solution at the microliter scale. The technique is based on the thermophoresis of molecules, which provides information about molecule size, charge, and hydration shell. Since at least one of these parameters is typically affected upon binding, the method can be used for the analysis of each kind of biomolecular interaction or modification of proteins or DNA. Quantitative binding parameters are obtained by using a serial dilution of the binding substrate. This section provides a detailed protocol describing the analysis of DNA–protein interactions, using the AT-hook peptides as a model system that bind to short double-stranded DNA.

Key words

Binding assay Dissociation constant DNA–protein interactions Microscale thermophoresis Interaction affinity 



The authors would like to thank Christoph J. Wienken the fruitful comments and suggestions for data analysis.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Universität Regensburg, Biochemistry IIIRegensburgGermany
  2. 2.NanoTemper Technologies GmbHMünichGermany
  3. 3.Ludwig-Maximilians-Universiät München, System BiophysicsMünichGermany
  4. 4.NanoTemper Technologies GmbHMünichGermany

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