JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB112155

Pyrophosphate Assay Kit (Fluorometric)

4

(2 Reviews)

|

(8 Publications)

Pyrophosphate Assay Kit (Fluorometric) ab112155 provides the most robust spectrophotometric method for the measurement of pyrophosphate.
6 Images
Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)
  • FuncS

Lab

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)

Pyrophospfate measured in cell lysates showing quantity (nmol) per 106cells

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)
  • FuncS

PubMed

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)

Urinary total pyrophosphate (PPi) concentrations were determined using ab112155. 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.

Caballero D et al., PLoS One, 12(7). Fig 1a. doi: 10.1371/journal.pone.0180098 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)
  • FuncS

Lab

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)

Pyrophosphate measured in tissue lysates showing quantity (nmol) per microgram protein

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)
  • FuncS

Supplier Data

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)

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.

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)
  • FuncS

Lab

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)

Example PPi standard curve.

Data provided for demonstration purposes only.

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)
  • FuncS

Lab

Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (AB112155)

Pyrophosphate measured in biofluids showing concentration (micromolar). Obtained values were recalculated from 1 : 1000 sample dilutions.

Key facts

Detection method

Fluorescent

Sample types

Tissue Lysate, Urine, Plasma, Cell culture extracts, Suspension cells, Serum, Adherent cells

Results type

Quantitative

Assay Platform

Microplate reader

style
left-column

Complementary Products

Secondary Antibodies

AB150077

Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)

Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (AB150077)

  • 5
  • 20
Assay Kits

AB65622

Phosphate Assay Kit (Colorimetric)

Functional Studies - Phosphate Assay Kit (Colorimetric) (AB65622)

  • 0
  • 0
Primary Antibodies

AB15580

Anti-Ki67 antibody

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (AB15580)

  • 5
  • 230
Sample Prep & Detection Kits

AB93349

10kDa Spin Column

Other - 10kDa Spin Column (AB93349)

  • 5
  • 1
style
left-column

Product details

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).

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

style
left-column

What's included?

{ "values": { "2x96Tests": { "sellingSize": "2 x 96 Tests", "publicAssetCode":"ab112155-2x96Tests", "assetComponentDetails": [ { "size":"1 x 25 mL", "name":"Assay Buffer", "number":"AB112155-CMP01", "productcode":"" }, { "size":"1 x 1 Vial", "name":"PPi Sensor", "number":"AB112155-CMP03", "productcode":"" }, { "size":"1 x 1 mL", "name":"Pyrophosphate Standard (50mM)", "number":"AB112155-CMP04", "productcode":"" }, { "size":"1 x 200 µL", "name":"DMSO", "number":"AB112155-CMP02", "productcode":"" } ] } } }
style
left-column

Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
style
left-column

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.
Biological function summary

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.

Pathways

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.
style
left-column

Product protocols

style
left-column

Target data

style
left-column

Publications (8)

Recent publications for all applications. Explore the full list and refine your search

Journal of bone and mineral metabolism 42:681-698 PubMed39212714

2024

Enpp1 mutations promote upregulation of hedgehog signaling in heterotopic ossification with aging.

Applications

Unspecified application

Species

Unspecified reactive species

Zhongyuan He,Zhengya Zhu,Tao Tang,Fuan Wang,Peng Guo,Jianfeng Li,Nguyen Tran Canh Tung,Qian Liang,Shaoyu Liu,ManMan Gao,Xizhe Liu,Zhiyu Zhou

eLife 13: PubMed38334260

2024

O-GlcNAc signaling increases neuron regeneration through one-carbon metabolism in .

Applications

Unspecified application

Species

Unspecified reactive species

Dilip Kumar Yadav,Andrew C Chang,Noa W F Grooms,Samuel H Chung,Christopher V Gabel

Bone 164:116525 PubMed35987514

2022

Phosphate promotes osteogenic differentiation through non-canonical Wnt signaling pathway in human mesenchymal stem cells.

Applications

Unspecified application

Species

Unspecified reactive species

Shumin Rui,Takuo Kubota,Yasuhisa Ohata,Kenichi Yamamoto,Makoto Fujiwara,Shinji Takeyari,Keiichi Ozono

Bone 105:134-147 PubMed28866368

2017

Overlapping functions of bone sialoprotein and pyrophosphate regulators in directing cementogenesis.

Applications

Unspecified application

Species

Unspecified reactive species

M Ao,M B Chavez,E Y Chu,K C Hemstreet,Y Yin,M C Yadav,J L Millán,L W Fisher,H A Goldberg,M J Somerman,B L Foster

PloS one 12:e0180098 PubMed28704395

2017

Intraperitoneal pyrophosphate treatment reduces renal calcifications in Npt2a null mice.

Applications

Unspecified application

Species

Unspecified reactive species

Daniel Caballero,Yuwen Li,Jonathan Fetene,Julian Ponsetto,Alyssa Chen,Chuanlong Zhu,Demetrios T Braddock,Clemens Bergwitz

EMBO reports 16:520-7 PubMed25648146

2015

The cAMP phosphodiesterase Prune localizes to the mitochondrial matrix and promotes mtDNA replication by stabilizing TFAM.

Applications

FuncS

Species

Unspecified reactive species

Fan Zhang,Yun Qi,Kiet Zhou,Guofeng Zhang,Kaari Linask,Hong Xu

Nature chemical biology 10:450-6 PubMed24776930

2014

Plant perception of β-aminobutyric acid is mediated by an aspartyl-tRNA synthetase.

Applications

Unspecified application

Species

Unspecified reactive species

Estrella Luna,Marieke van Hulten,Yuhua Zhang,Oliver Berkowitz,Ana López,Pierre Pétriacq,Matthew A Sellwood,Beining Chen,Mike Burrell,Allison van de Meene,Corné M J Pieterse,Victor Flors,Jurriaan Ton

Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 28:1135-49 PubMed23184610

2012

Loss of equilibrative nucleoside transporter 1 in mice leads to progressive ectopic mineralization of spinal tissues resembling diffuse idiopathic skeletal hyperostosis in humans.

Applications

Unspecified application

Species

Unspecified reactive species

Sumeeta Warraich,Derek B J Bone,Diana Quinonez,Hisataka Ii,Doo-Sup Choi,David W Holdsworth,Maria Drangova,S Jeffrey Dixon,Cheryle A Séguin,James R Hammond
View all publications
style
left-column
style
left-column
websiteProtocolBooklet
en
style
right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com