Table 1. Comparison of the AV-BcO association rate constants (k_on) obtained by fluorescence change, $\overline{\mathit{F}}\left(\mathit{t}\right)$, and corrected fluorescence anisotropy, $\overline{\mathit{r}\mathit{F}}\left(\mathit{t}\right)$.

kon AV-BcO	$\overline{\mathbf{F}}\left(\mathbf{t}\right)$ × 10−6 M-1s-1			$\overline{\mathbf{r}}\mathbf{F}\left(\mathbf{t}\right)$ × 10−6 M-1s-1
Temp.	260 nM	520 nM	1040 nM	260 nM	520 nM	1040 nM
25°C	9.5 ± 0.1	9.5 ± 0.3	9.7 ± 0.3	9.5 ± 0.1	9.5 ± 0.1	9.4 ± 0.2
20°C	5.7 ± 0.1	5.9 ± 0.3	6.0 ± 0.1	5.9 ± 0.1	5.8 ± 0.1	6.1 ± 0.1
15°C	4.1 ± 0.1	4.0 ± 0.1	3.9 ± 0.2	4.0 ± 0.1	3.8 ± 0.1	4.0 ± 0.2
10°C	2.4 ± 0.1	2.6 ± 0.2	2.5 ± 0.2	2.7 ± 0.1	2.7 ± 0.1	2.7 ± 0.1

The association reactions were acquired with BcO (20 nM) binding to AV at several temperatures, protein concentrations and pH 8.The $\overline{F}\left(t\right)$ and $\overline{rF}\left(t\right)$ signals were equivalent as they tracked in the errors the association process of dye-labeled B₇ binding to the proteins under pseudo-first order conditions.