where F0 and F correspond to the fluorescence emission of the
protein in the absence and presence of quencher at a given
concentration [Q]. The slope of the plot F0F over [Q] is the
Stern–Volmer constant (KSV), which is defined by the product of
the bimolecular rate constant of the quenching process (kq) and
the fluorophore lifetime in the absence of quencher () (36).
Because acrylamide absorbs weakly at 295 nm, fluorescence
intensity was corrected as described (37).