| Assay Method Information | |
| | ITC Assay |
| Description: | All ITC experiments were performed using a VP-ITC (Microcal, Northampton, MA) instrument at 10 or 15 °C in 50 mM HEPES, pH 7.5, 300 mM NaCl, and 1 mM DTT, and all protein samples were dialyzed against this buffer in advance. The concentration of the proteins was between 0.1 mM and 0.2 mM, while the ligand was 10 to 15 times less. A typical titration was conducted using an initial injection of 2 μL, followed by 25 identical injections of 10 μL at time intervals of at least 240 s. |
| Affinity data for this assay | |
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