Fig. 1.

Simulated saturation and kinetic binding curves. Saturation and kinetic binding curves were simulated in GraphPad prism assuming R + L ⇋ RL using the one site: total and non-specific binding equation (A) or association then dissociation equation (B) for a ligand with a  K_d of 5nM (A) or 50nM (B). For (A), the specific binding (squares) B_max was set to 100 and K_d to 5 nM and the non-specific binding (triangles) set to a slope of 0.25. For (B) the k_on was set to 1 × 10⁶ M⁻¹ min⁻¹ and k_off to 0.05 min⁻¹, the concentration of ligand to 50 nM and the B_MAX to 150, which results in a K_d of 50 nM. The association phase of the kinetic binding curve is termed k_on(obs) and is defined as k_on = (k_on(obs)-k_off)/[ligand].