Pyrophosphate (PPi) Assay Kit(Fluorometric/Colorimetric)

AgK-379

Pyrophosphate (PPi) Assay Kit(Fluorometric/Colorimetric)

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 (Ex/Em = 535/587 nm) or color (OD: 570 nm) intensities are directly proportional to the concentrations of pyrophosphate, enabling precise measurements. Monomeric inorganic phosphate (Pi) does not interfere with the assay. Our assay delivers an easy and robust method suitable for use in a variety of biological samples and can be performed in a convenient microtiter-plate format. The kit provides sufficient reagents for 100 fluorometric or 50 colorimetric assays, respectively. This kit can detect as low as 1.8 μM PPi in plasma and serum samples.

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.

• Determination of PPi concentration in biological samples
• Monitor pyrophosphate release by a variety of enzymes
• Screening inhibition or activities of enzymes that consume pyrophosphate

Store at -20°C.

100 assays

• PPi Assay Buffer
• PPi Buffer Supplement
• PPi Substrate
• PPi Enzyme Mix
• PPi Developer
• PPi Probe
• PPi Standard (1 mM)

Plate Reader capable of 37°C temperature setting and readings of fluorescence (Ex/Em 535/587 nm) or absorbance (570 nm)
96-well clear plates with flat bottom
10 kDa spin columns for sample preparation

Detection method- Fluorescence (Ex/Em = 535/587 nm) or Absorbance (OD 570 nm)

Plasma, serum and other biological fluids
Cell culture extracts and tissue lysates