Fig 5. Concentration dependence of association traces of BcO binding to AV and BFl binding to SAV.

(A) Corrected Fluorescence Anisotropy Concentration dependence of BcO (20 nM) binding to AV at pH 8 and 20°C, normalized as $\overline{rF}\left(t\right)=\left[rF\left(0\right)-rF\left(t\right)\right]/\left[rF\left(0\right)-rF\left(\infty \right)\right]$. The observed (color) curves were obtained with an AV concentration of 1040 nM (top), 520 nM (middle) and 260 nM (bottom) and the fitted curves (black lines) had halftimes of 109 ms, 229 ms and 455 ms; respectively. (B) Shows the corresponding semi-logarithmic plots of $\overline{rF}\left(t\right)$. (C) Normalized fluorescence change, $\overline{F}\left(t\right)$, concentration dependence of BFl (20 nM) binding to SAV at pH 8 and 20°C. Normalized as followed: $\overline{F}\left(t\right)=\frac{F\left(t\right)}{F\left(0\right)}=\left[F\left(t\right)-rF\left(\infty \right)\right]/\left[rF\left(0\right)-rF\left(\infty \right)\right]$. The observed (color) curves were acquired with SAV concentrations of 200 nM (red), 400 nM (purple) and 800 nM (orange) where the fitted curves (black lines) had halftimes of 77.3 ms, 37.7 ms and 20.6 ms; respectively. (D) Shows the corresponding semi-logarithmic plot of $\overline{F}\left(t\right)$.