| Assay Method Information | |
| | Isothermal Titration Calorimetry |
| Description: | ITC experiments were performed at 25° C. using a MicroCal PEAQ-ITC microcalorimeter (Malvern). PAH proteins were prepared in a HEPES buffer (HEPES 20 mM Hepes, 0.2M NaCl, 1 mM DTT, pH 7.0) containing 5% DMSO, in order to match the final DMSO concentration of the compounds dilutions. The protein concentration used (30 μM) was determined using ultraviolet (UV) absorbance at 280 nm. The compounds (500 mM) were prepared using the exact same buffer. Twelve injections of 3 μL of compound/ligand were titrated into the sample cell (containing the protein) over 31 min with a stirring speed of 750 rpm. Data was baseline adjusted by subtracting the background obtained from equivalent injections of compound into the buffer solution. |
| Affinity data for this assay | |
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