For high-throughput inhibitor screening and evaluation of pyrophosphatase modulators.

• Safe and non-radioactive assay
• Fast and convenient
• The procedure involves incubation of enzyme and inhibitor, addition of a single working reagent and incubation for 30 min
• Room temperature assay
• No 37 °C incubator is needed
• High-throughput
• Homogenous 'mix-incubate-measure' type assay
• Can be readily automated on HTS liquid handling systems
• Robust assay with a Z' factor of 0.59
• Can be used in 96-well, 384-well, and potentially higher density screening assays

Inorganic pyrophosphatase (E.C.3.6.1.1) catalyzes the hydrolysis of phosphoester bonds of inorganic pyrophosphate [P2O74-], thereby releasing two orthophosphate molecules. Family I PPases are essential enzymes found in all kingdoms of life and are responsible for maintaining the correct pyrophosphate equilibrium necessary to carry out nucleic acid and protein synthesis, and facilitate fatty acid beta oxidation.