Figure 2

Optimization of BCS-Cu+ binding assay. (A) Schematic depiction of BCS binding with cuprous ion. (B) Absorption spectra of Cu+, BCS and Cu+-BCS. (C) The effect of ascorbate on BCS assay (50 mM Tris–HCl pH 7.4, 200 µM BCS, 25 μM CuSO4). (D) The effect of BCS on BCS assay (50 mM Tris–HCl pH 7.4, 25 μM CuSO4, 1 mM ascorbate). (E) The effect of CuSO4 on BCS assay (50 mM Tris–HCl pH 7.4, 200 µM BCS, 1 mM ascorbate). (F) The effect of pH on BCS assay (200 µM BCS, 25 μM CuSO4, 1 mM ascorbate). All assays were conducted at least 6 times. Average ± S.D. is presented. Asterisk * indicates significant difference (* p < 0.05, **p < 0.01).