Kinetics of non-labeled interactors
| To determine on and off rates of non-labeled interactors, we developed an in-house software which leverages the model developed by Motulsky and Mahan in 1983. In that framework, the authors derived equations that allow determination of on and off rates of non-labeled ligands upon mixture with target and labeled ligand. As input, our algorithm requires the on- and off- rates of the labeled ligand interaction in addition to the concentrations of labeled and non-labeled ligands. Affinities calculated based on the reaction rate constants are always in line with those calculated from equilibrium experiments. Moreover, we have recently established jump dilutions to examine kinetics for as long as 50 hours. |