Pyrophosphate Assay Kit (Fluorometric) ab112155 provides the most robust spectrophotometric method for the measurement of pyrophosphate.
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Pyrophosphate Assay Kit (Fluorometric) ab112155 provides the most robust spectrophotometric method for the measurement of pyrophosphate.
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) (Pyrophosphate Assay Kit II (Fluorometric) ab179836).
Other Notes
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.
Samples such as plasma and serum might need a deproteinization step with spin columns
Inorganic pyrophosphate often abbreviated as PPi acts as a byproduct of several biosynthetic reactions. It possesses a molecular mass of 177.98 g/mol. Pyrophosphate functions as an energy-rich intermediate that participates actively in reactions like the synthesis of nucleic acids and proteins. Inorganic pyrophosphatase an enzyme frequently associated with pyrophosphate metabolism catalyzes the hydrolysis of PPi releasing inorganic phosphate. This hydrolysis occurs release energy facilitating various biochemical processes. Pyrophosphate molecules are detected in tissues with high cell proliferation such as liver kidney and bones.
Inorganic pyrophosphate plays a critical part in bone mineralization and the regulation of calcium and phosphate metabolism. It influences the bone matrix's deposition by calcifying or inhibiting mineralization depending on the balance with phosphates. Within the extracellular matrix pyrophosphate remains associated with proteins such as alkaline phosphatase which converts PPi to phosphate promoting mineralization. Pyrophosphate's activity shows strong relevance in maintaining extracellular fluid stability preventing excessive calcification.
The involvement of inorganic pyrophosphate occurs prominently in the regulation of the ATP metabolism pathway and cartilage formation. It is closely related to the enzyme family of pyrophosphatases which include both soluble inorganic pyrophosphatase and membrane-integrated soluble pyrophosphatases. These enzymes orchestrate converting pyrophosphate into orthophosphate ions feeding into the pathways that regulate energy homeostasis and biomineralization. Interaction with proteins like ENPP1 and TNAP highlight its role within the broader context of nucleic acid synthesis and tissue calcification.
Abnormal levels of inorganic pyrophosphate link to skeletal disorders like osteoporosis and osteoarthritis. High pyrophosphate concentrations can lead to pathological calcification processes prominently affecting joint health and bone density. The enzyme ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) connects to pyrophosphate's regulation of these diseases where it influences extracellular pyrophosphate concentration and subsequently mineralization. Understanding pyrophosphate dynamics provides insights into the pathophysiology of metabolic bone diseases.
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Terms & Conditions.
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
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.
Pyrophosphate measured in biofluids showing concentration (micromolar). Obtained values were recalculated from 1:1000 sample dilutions.
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