I want to understand the general principle while establishing a competitive assay. Should it be between a binding site vs two ligands (hot and cold peptides in case of peptide) competing for it or it can be either binding site (protein) or ligands in two forms vs the other interacting partner?
Adam B Shapiro
In theory, there is no distinction between the ligand (often a small molecule) and the receptor (often a macromolecule) in binding equations. They are interchangeable. So, you can use 2 small ligands competing for binding to a single protein, or 2 proteins competing for binding to a single small ligand.
In practice, I think it is more common to use two ligands and one protein.
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