| Assay Method Information | |
| | Isothermal Titration Calorimetry (ITC) Measurements |
| Description: | All calorimetric experiments were carried out using an Affinity ITC from TA Instruments (New Castle, Del.) equipped with an auto sampler in a buffer containing 20 mM HEPES, pH 7.5, 150 mM NaCl, 0.5 mM TCEP, and 2% DMSO at 25° C. 20 μM protein solution containing PRPK or Cereblon (CRBN) in the calorimetric cell was titrated with 200 μM of a test compound solution using 2 μL injection in 200 seconds intervals using a stirring speed at 125 rpm. Resulting isotherm was fitted with a single site model to yield thermodynamic parameters of ΔH, ΔS, stoichiometry, and Kd using NanoAnalyze software (TA instruments). |
| Affinity data for this assay | |
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