(A) Standard curve generated using known amounts of Pi from sodium phosphate dissolved in ultrapure water combined with the ammonium molybdate/malachite green reagent as described in materials and methods. Color was allowed to develop for 10 minutes at room temperature before measuring A660. (B) Simultaneous assay measurement of 45Ca2+ accumulation (left y-axis) and Pi release (right y-axis) in the presence of 300 nM free Ca2+. Reactions were carried out with 3 mM Mg2+, 2 mM ATP for 10 minutes at 37°C with no additional treatments (control) or in the presence of 100 nM TG or 10 μM Ca2+ ionophore A23187.
McMullen et al. Biological Procedures Online 2012 14:4 doi:10.1186/1480-9222-14-4