Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Sample type: Adherent cells, Cell culture extracts, Plasma, Serum, Suspension cells, Tissue Lysate, Urine
Product namePyrophosphate Assay Kit (Fluorometric)
See all Pyrophosphate kits
Sample typeUrine, Serum, Plasma, Cell culture extracts, Adherent cells, Suspension cells, Tissue Lysate
Pyrophosphate Assay Kit (Fluorometric) ab112155 provides the most robust spectrophotometric method for the measurement of pyrophosphate.
The pyrophosphate assay protocol uses a proprietary fluorogenic pyrophosphate sensor that has its fluorescence intensity proportionally dependent upon the concentration of pyrophosphate.
This pyrophosphate assay protocol is much easier and more robust than enzyme-coupling pyrophosphate methods, which require at least two enzymes for their pyrophosphate detections. Due to its direct measurement of pyrophosphate, this kit is ideal for screening inhibition or activities of enzymes that consume or generate pyrophosphate.
The pyrophosphate assay is an optimized mix-and-read assay and can be performed in a convenient 96-well or 384-well microtiter-plate format.
Assay time: 10-30 minutes
This product measures PPi formation at Ex/Em = 316/456 nm.
If your instrument has filters for Ex/Em = 370/470 nm, we recommend using Pyrophosphate Assay Kit II (Fluorometric) (ab179836).
Pyrophosphates (PPi) are produced by a number of biochemical reactions, such as ATP hydrolysis, 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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 2 x 96 tests Assay Buffer 1 x 25ml DMSO 1 x 200µl PPi Sensor 1 vial Pyrophosphate Standard (50mM) 1 x 1ml
Urine PPi was corrected for urine creatinine, which was measured by LC-MS/MS or by ELISA using appropriate controls to adjust for inter-assay variability.
Pyrophospfate measured in cell lysates showing quantity (nmol) per 106cells
Pyrophosphate measured in tissue lysates showing quantity (nmol) per microgram protein
Pyrophosphate measured in biofluids showing concentration (micromolar). Obtained values were recalculated from 1:1000 sample dilutions.
Example PPi standard curve.
Data provided for demonstration purposes only.
Pyrophosphate, ATP and phosphate dose responses were measured with ab112155 in a solid black 96-well plate using a fluorescence microplate reader. As low as 1 µM (100 picomoles/well) pyrophosphate can be detected with 10 minutes incubation.
ab112155 has been referenced in 4 publications.
- Caballero D et al. Intraperitoneal pyrophosphate treatment reduces renal calcifications in Npt2a null mice. PLoS One 12:e0180098 (2017). PubMed: 28704395
- Ao M et al. Overlapping functions of bone sialoprotein and pyrophosphate regulators in directing cementogenesis. Bone 105:134-147 (2017). PubMed: 28866368
- Zhang F et al. The cAMP phosphodiesterase Prune localizes to the mitochondrial matrix and promotes mtDNA replication by stabilizing TFAM. EMBO Rep 16:520-7 (2015). Functional Studies . PubMed: 25648146
- Luna E et al. Plant perception of ß-aminobutyric acid is mediated by an aspartyl-tRNA synthetase. Nat Chem Biol 10:450-6 (2014). PubMed: 24776930