A protein binding-induced supramolecular dissociation strategy is developed with the ratio of monomer and excimer fluorescence as the tool for protein sensing and quantification. Due to the "lock-and-key" strategy based on specific ligand-protein binding, the probe exhibits excellent selectivity and quantification accuracy to the protein of interest. The ratiometric approach is immune to interference from extrinsic quenchers, while preserving the opportunity to be protein specific.