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Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040) provides a fast and convenient method for determination of free inorganic pyrophosphate (PPi) levels in biological material.

Pyrophosphate Assay Kit (Colorimetric / Fluorometric) designed for success:
- Can be used with a variety of sample types, including tissue extracts and body fluids
- Highly sensitive, of which the detection limit can be as low as 1.8 µM PPi in plasma and serum samples
- Can be perfomed using either fluorescence or colorimetric detection

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Images

Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (AB234040), expandable thumbnail
  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (AB234040), expandable thumbnail
  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (AB234040), expandable thumbnail
  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (AB234040), expandable thumbnail
  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (AB234040), expandable thumbnail

Publications

Key facts

Detection method
Colorimetric/Fluorometric
Sample types
Tissue Lysate, Plasma, Cell culture extracts, Serum, Other biological fluids
Assay type
Quantitative

Associated Products

Select an associated product type

1 product for Alternative Product

What's included?

100 Test
Components
AMP
1 x 200 µL
Assay Buffer 15
1 x 25 mL
Developer Mix A
1 x 1 Vial
Enzyme Mix X
1 x 200 µL
OxiRed™ Probe
1 x 0.2 mL
PEP
2 x 1 Vial
Pyrophosphate Standard
1 x 200 µL

Recommended products

Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040) provides a fast and convenient method for determination of free inorganic pyrophosphate (PPi) levels in biological material.

Pyrophosphate Assay Kit (Colorimetric / Fluorometric) designed for success:
- Can be used with a variety of sample types, including tissue extracts and body fluids
- Highly sensitive, of which the detection limit can be as low as 1.8 µM PPi in plasma and serum samples
- Can be perfomed using either fluorescence or colorimetric detection

Key facts

Detection method
Colorimetric/Fluorometric
Sample types
Tissue Lysate, Plasma, Cell culture extracts, Serum, Other biological fluids
Assay type
Quantitative
Assay Platform
Microplate reader

Storage

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

Notes

How the assay works

In Pyrophosphate Assay Kit (Colorimetric / Fluorometric)assay protocol, 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.

Pyrophosphate Assay Kit (Colorimetric / Fluorometric)protocol summary:

- Prepare samples.
- Prepare standard curve.
- Prepare reaction mix.
- Add samples, controls and standards to wells and incubate plate protected from light.
- Measure absorbance (570 nm) or fluorescence EX/Em = 535/587 nm).

Related and recommended products

See an alternative Pyrophosphate Assay Kit:
Pyrophosphate Assay Kit (Fluorometric) Pyrophosphate Assay Kit (Fluorometric) ab112155

This product is manufactured by BioVision, an Abcam company and was previously called K568 Pyrophosphate (PPi) Assay Kit (Fluorometric/Colorimetric). K568-100 is the same size as the 100 test size of ab234040.

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

Supplementary info

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

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040), expandable thumbnail

    Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040)

  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040), expandable thumbnail

    Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040)

    Pyrophosphate measured in HepG2 and HeLa cell lysates.

    Cells were extracted directly in the Assay Buffer.

  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040), expandable thumbnail

    Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040)

    Spike and recovery in normal human plasma.

    Spike and recovery in 20 μL of normal human plasma. Plasma samples were spiked with 0.06 nmol of PPi Standard and assayed according to kit protocol yielding 96% PPi recovery.

  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040), expandable thumbnail

    Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040)

  • Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040), expandable thumbnail

    Functional Studies - Pyrophosphate Assay Kit (Colorimetric / Fluorometric) (ab234040)

    Quantification of pyrophosphate in de-proteinized undiluted human serum and pooled plasma.

    Quantification of pyrophosphate in 20 μL of de-proteinized undiluted human serum and pooled plasma.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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

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