Invitrogen™ EnzChek™ Pyrophosphate Assay Kit

Invitrogen™ EnzChek™ Pyrophosphate Assay Kit

The EnzChek™ Pyrophosphate Assay Kit provides a rapid and highly sensitive enzymatic assay for detecting free pyrophosphate in solution. In this assay the formation of a chromophoric product is detected spectrophotometrically.

The EnzChek™ Pyrophosphate Detection Kit provides a fast, convenient, and inexpensive spectrophotometric method for measuring the inorganic pyrophosphate produced by a number of biochemical reactions, such as DNA and RNA polymerizations, cyclic AMP formation by the enzyme adenylate cyclase, and the enzymatic activation of fatty acids to form their coenzyme A esters.
The EnzChek™ Pyrophosphate Assay Kit is a modification of our EnzChek™ Phosphate Assay Kit (Cat. No. E6646). In the presence of inorganic phosphate, the substrate 2-amino-6-mercapto-7-methylpurine ribonucleoside (MESG) is converted by the purine nucleoside phosphorylase (PNP) enzyme to ribose 1-phosphate and 2-amino-6-merc apto-7-methylpurine. The enzymatic conversion of MESG results in a shift in absorbance maximum from 330 nm to 360 nm. The EnzChek™ Pyrophosphate Assay Kit includes the enzyme inorganic pyrophosphatase, which catalyzes conversion of pyrophosphate into two equivalents of phosphate. The phosphate is then consumed by the MESG/PNP reaction and detected by an increase in absorbance at 360 nm. Additional sensitivity is gained by the amplification of one molecule of pyrophosphate into two molecules of phosphate.

Features

• Sensitivity limit of 1 µM pyrophosphate (∼0.2 µg/mL)
• Acommodates a pH range of 6.5 to 8.5

Contents

• Includes inorganic pyrophosphatase, 2-Amino-6-mercapto-7-methylpurine riboside (MESG), purine nucleoside phosphorylase (PNP), concentrated reaction buffer, Na2P2O7 standard, and detailed protocols for detecting and quantitating pyrophosphate.