The EnzChek® Phosphate Assay Kit provides a fast and sensitive spectrophotometric method for the quantification of inorganic phosphate in solution and for phosphate released during ATPase or GTPase enzymatic reactions.
• Sensitivity of 2 to 150 µM phosphate • Spectrophotometric microplate-based assay detects a change in absorbance maximum from 330 nm to 360 nm • Continuous monitoring of phosphate generated by ATPase or GTPase activity
In the presence of inorganic phosphate, the 2-amino-6-mercapto-7-methylpurine riboside (MESG) substrate is converted by the purine nucleoside phosphorylase (PNP) enzyme to the ribose 1-phosphate and 2-amino-6-mercapto-7-methylpurine product. This enzymatic conversion of MESG results in a spectrophotometric shift in maximum absorbance from 330 nm for the substrate to 360 nm for the product.
Sensitivity of the assay is in the range of 2 to 150 µM inorganic phosphate (2 to 150 nanomoles phosphate in a 1 mL volume), and the reaction can be performed over a pH range of 6.5 to 8.5. The EnzChek® phosphate reaction is sufficiently fast and quantitative for use in stopped-flow kinetic experiments.
For Research Use Only. Not for use in diagnostic procedures.