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Isothermal titration calorimetry

HG Hannah Girstmair
FT Franziska Tippel
AL Abraham Lopez
KT Katarzyna Tych
FS Frank Stein
PH Per Haberkant
PS Philipp Werner Norbert Schmid
DH Dominic Helm
MR Matthias Rief
MS Michael Sattler
JB Johannes Buchner
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To determine the binding affinity of the Hsp90 NTDs to ATP and RD ITC was applied. Prior to the measurements the Hsp90 NTDs were dialyzed against ITC buffer (40 mM HEPES, pH 7.5, 150 mM KCl, 5 mM MgCl2). The same buffer was used to prepare the ligand stock solution. The concentration of the Hsp90 NTD Stock solution was 10 µM and the ATP and RD stock concentrations were 100 µM. The measurements were performed with a MicroCal PEAQ-ITC calorimeter (Malvern Instruments Limited, UK). All ITC measurements were performed at 25 °C. Data analysis was carried out with the MicroCal user software.

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