Assay Method Information

Assay Name:  Enzyme Inhibition Assay
Description:  An enzyme assay method using isothermal titration calorimetry (ITC) was developed to investigate the inhibition kinetics of ODCase. Titrations were performed on an isothermal titration calorimeter, VP-ITC instrument from MicroCal (Northampton, MA). In all cases, the calorimetric cell contained the enzyme solution while the ligand(s) were loaded into the automatic injection syringe. Substrate and inhibitor mixtures were prepared and loaded into the injection syringe to monitor enzyme activity in the presence of inhibitor. Single injection method was used in all studies. The heat change was measured as the reaction progressed until the heat returned to baseline, indicating the end of the reaction. The inhibition constant, Ki was calculated by direct fitting of the data from inhibition assays, or was derived from the Dixon plot for competitive inhibition.
Affinity data for this assay
 

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