Pyrophosphate (PPi) Assay Kit (Fluorometric/Colorimetric)

Inorganic pyrophosphate (diphosphate, pyrophosphoric acid or PPi) is a small molecule formed during, or required as a substrate, for a number of biochemical reactions. Enzymatic reactions that generate PPi include ATP hydrolysis, DNA and RNA polymerization, cyclic AMP formation, and enzymatic activation of fatty acids to form their CoA esters. Conversely, enzymes that utilize PPi may include cyclases, hydrolases and ligases. Measurement of inorganic pyrophosphate (PPi) in plasma, serum and other biologic fluids is important in studies of bone metabolism, renal stone disease, and certain types of arthritis.

The kit is suitable for the determination of PPi concentration, monitoring pyrophosphate release by a variety of enzymes and for the screening inhibition or activities of enzymes that consume pyrophosphate. Suitable samples include plasma, serum and other biological fluids, cell culture extracts and tissue lysates.

The Pyrophosphate (PPi) Assay Kit provides a fast, convenient and ultrasensitive method for determination of free inorganic pyrophosphate levels in biological material. PPi produced during biotic processes is detected through a series of reactions which utilize a proprietary enzyme mix and probe, generating a stable product that can be quantified by either colorimetric or fluorometric readout. Generated fluorescence (λ_Ex = 535 nm/λ_Em = 587 nm) or color (A₅₇₀ nm) intensities are directly proportional to the concentrations of pyrophosphate, enabling precise measurements. Monomeric inorganic phosphate (Pi) does not interfere with the assay.

The Pyrophosphate (PPi) Assay Kit is:

• Compatible with high-throughput handling systems.
• Specific for Pyrophosphate: other molecules containing phosphate groups do not interfere with the assay.
• Detects as little as 2 μM of Pyrophosphate in a variety of biological samples.