| Assay Method Information | |
| | ITC Titration |
| Description: | ITC experiments were carried out using an iTC200 microcalorimeter (MicroCal) with coin shaped sample cell (200 μL) at 37 C with stirring at 1000 rpm. Depending on the solubility of the samples, protein was placed in the cell or the syringe for the titration. Compounds that were readily solubilized (ANS, ketorolac and fenofibric acid) were diluted from concentrated stocks in identical buffer (20 mM HEPES pH 8, 50 mM NaCl, 0.5 mM EDTA), and placed in the titration syringe and hIFABP was place in the sample cell. In the case of compounds with limited solubility (DAUDA, GW7647 and L165041), hIFABP was placed in the titration syringe and concentrated stocks of the compounds in DMSO were diluted in identical buffer and placed in the sample cell while maintaining 0.5% of DMSO solvent. Typically, 16 serial injections at intervals of 220 s were made with continuous stirring of the solution in the sample cell |
| Affinity data for this assay | |
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