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

2011; Springer Science+Business Media; Linguagem: Inglês

10.1007/978-1-61779-424-7_18

1940-6029

Karina Zillner, Moran Jerabek‐Willemsen, Stefan Duhr, Dieter Braun, Gernot Längst, Philipp Baaske,

thermodynamics and calorimetric analyses

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.