Inorganic pyrophosphate (also referred to as diphosphate, pyrophosphoric acid or PPi) is a small diphosphate molecule that is required as a substrate or is the product formed from a number of different enzymatic reactions. Enzymes that utilize PPi as a substrate may include phosphotransferases (EC 2.7) and pyrophosphatases (EC 3.6.1.1).

• Fast - Measure enzyme activity via pyrophosphate consumption in 1 hour
• Simple - The 2 step, luminescent assay doesn’t require radioactive substrates, beads, detection antibodies or modified substrates
• Wide Detection Range - Sensitive to 0.02 µM with a linear range to 10 µM
• Versatile - Scalable to 96-, 384-, 1586-well formats

Enzymes that generate PPi are more numerous and may include cyclases (EC 4.6.1), hydrolyses (EC 3), ligases (EC 6) and DNA polymerases such as DNA Polymerase theta (θ) encoded by POLQ.

The PPiLight® Assay is a non-radioactive bioluminescent assay for the detection of inorganic pyrophosphate. In the presence of PPi the detection reagent catalyses the conversion of AMP to ATP. The assay uses luciferase, which produces light from the newly formed ATP and luciferin.

Measures activity of: Phosphotransferases, Pyrophosphotases, Cyclases, Ligases, Hydrolases, DNA Polymerases.

Storage Conditions: 2 - 8 °C.