Figure 6.

The K_D of HYlight for fructose 1,6-bisphosphate (FBP) is ∼1 μM, is not changing in the physiological pH range, and is affected by phosphorous compounds and changes in ionic strength

(A) Relative change of the fluorescence ratio as a function of increasing FBP concentrations and different buffers.

(B) Relative change of the fluorescence ratio as a function of increasing FBP concentrations and pH variation with the MES/CHES/HEPES buffer.

(C) Comparison of the ratio change in HYlight and HYlight Null in PBS, and TBS without FBP.

(D) Comparison of the ratio change in HYlight in the presence of 1 μM FBP and increasing NaCl concentrations.

(E) Relative change of the fluorescence ratio as a function of increasing FBP concentrations and increasing concentrations of NaCl. ns: Not significant; ∗<0.05; ∗∗<0.005; ∗∗∗<0.0005; ∗∗∗∗<0.00005. Ordinary One-Way ANOVA Šídák's Multiple Comparisons Test. PBS: 137 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 1.8 mM KH₂PO₄; TBS: 150 mM NaCl, 50 mM Tris-HCl, pH 7.6; MES/CHES/HEPES 25 mM; Britton-Robinson phosphate: acid mixture of NaOH, CH₃COOH, H₃PO₄, H₃BO₃, KCl. Data are presented as the mean ± SD. Sample size is n = 3 in all cases. See also Figure S4 for additional details.