The Non-Radioactive Phosphatase Assay Systems determine the amount of free phosphate generated in a reaction by measuring the absorbance of a molybdate:malachite green:phosphate complex. They allow use of a variety of buffer conditions and substrates, including naturally phosphorylated proteins or synthetic phosphopeptides.
The effective range for the detection of phosphate released during an assay using the Phosphatase Assay Systems is 100–4,000pmol of phosphate.
Use with Purified Samples or Crude Cell and Tissue Extracts
In addition to measuring phosphatase activity in partially fractionated and purified samples, the Phosphatase Assay Systems can measure phosphatase activity in crude cell or tissue extracts. For this application, the high concentration of phosphate in these preparations is eliminated prior to performing the assay using the supplied Spin Columns, which rapidly and effectively remove free phosphate and other low-molecular-weight inhibitors from the sample. In addition, a unique Molybdate Dye Additive that is combined with the Molybdate Dye Solution aids in the solubilization of proteins exposed to the acid conditions of the Molybdate Dye Solution, which alone could potentially cause precipitation of the proteins.