The “Binding Affinity” template found in the “Assay Development” section of the DI.Control software can be used to quantify any molecular interaction between a fluorescently labeled Target molecule and one corresponding Ligand molecule. Usually, before measuring a binding affinity experiment, a Binding check experiment is done to establish the assay conditions and to determine whether an interaction can be measured. After this, a Binding Affinity experiment is performed to monitor the target's TRIC or Spectral Shift signals over a dilution series of the ligand to derive a binding affinity (Kd or EC50).

How to do a Binding Affinity experiment:

After preparing the dose-response with the ligand and target in the desired wells, one would create a new experiment. In the main menu select “Binding Affinity” from the “Assay Development” menu.

If you only click with the mouse on the first well that you want to begin the experiment, the system automatically selects the 12 horizontal wells to the left (A1…A12, i.e.). If one wants to select other orientations or a different number of assay points, keep the mouse still clicked and move in the desired direction and the number of wells one desires. Once the desired wells are selected, release the click button.