Overview

  • Product name
    Pyrophosphate Assay Kit (Fluorometric)
    See all Pyrophosphate kits
  • Sample type
    Urine, Serum, Plasma, Cell culture extracts, Adherent cells, Suspension cells, Tissue Lysate
  • Assay type
    Quantitative
  • Product overview

    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.

    Pyrophosphate Assay Kit (Fluorometric) provides the most robust spectrophotometric method for the measurement of pyrophosphate. It uses a proprietary fluorogenic pyrophosphate sensor that has its fluorescence intensity proportionally dependent upon the concentration of pyrophosphate. Abcam's assay 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 assay is an optimized mix-and-read assay and can be performed in a convenient 96-well or 384-well microtiter-plate format. The kit provides all the essential components for assaying pyrophosphate.

    Visit our FAQs page for tips and troubleshooting.

  • Notes

    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).

Properties

Images

  • Urinary total pyrophosphate (PPi) concentrations were determined using ab112155
    Urinary total pyrophosphate (PPi) concentrations were determined using ab112155Caballero D et al., PLoS One, 12(7). Fig 1a. doi: 10.1371/journal.pone.0180098

    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.

  • Functional Studies- Pyrophosphate Assay Kit (Fluorometric) (ab112155)
    Functional Studies- Pyrophosphate Assay Kit (Fluorometric) (ab112155)

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

  • Functional Studies- Pyrophosphate Assay Kit (Fluorometric) (ab112155)
    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)
    Functional Studies- Pyrophosphate Assay Kit (Fluorometric) (ab112155)

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

  • Functional Studies- Pyrophosphate Assay Kit (Fluorometric) (ab112155)
    Functional Studies- Pyrophosphate Assay Kit (Fluorometric) (ab112155)
  • Functional Studies - Pyrophosphate Assay Kit (Fluorometric) (ab112155)
    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.

Protocols

References

This product has been referenced in:
  • Caballero D  et al. Intraperitoneal pyrophosphate treatment reduces renal calcifications in Npt2a null mice. PLoS One 12:e0180098 (2017). Read more (PubMed: 28704395) »
  • 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 . Read more (PubMed: 25648146) »

See all 2 Publications for this product

Publishing research using ab112155? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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30 reviews or Q&A

10
Votes: Highest First

Hi, I have some technical questions about the product 
Pyrophosphate Assay Kit II (Fluorometric) (ab179836)
 

 

1) The closest wavelength on my instrument is Ex/Em=355/460 rather than the recommended Ex/Em=370/470 of this prod...

Read More

Thank you for contacting us.

To answer your questions:

1) Yes you can use ab179836 with the settings on your instruments

2) Ab112155 would not work for you, excitation 355 is too high for this kit

3) Yes, your sample...

Read More

Thank you very much for your standard curve. It's very helpful.

Thank you for your kind response. It is a pleasure to help in any way I can. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Measuring PPi in mouse serum

 Average Average 3/5 (Ease of Use)
Abreviews
We wanted to measure PPi in mouse blood. A big problem is the high autofluorescence of serum and plasma, possibly contributed by albumin. Therefore we tested several conditions:
- untreated serum
- heat-inactivated serum (45 min 55°C)
- plasma
- serum after centrifugation through a 300 kDa cut-off-membrane
PPi amount was expected in a range between 0.5 - 5 µM [Koba et al. 2005]. Only the last condition (cut-off-membrane) resulted in the expected measurement (approximately 1.62 µM).
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Verified customer

Submitted Jan 19 2017

RIPA buffer interfere with assay?

I can confirm that SDS, Tween-20, and EDTA (> 30 um) will interfere with the assay. Since RIPA buffer has SDS in it, it is not suitable.

TritonX-100 up to 0.5% does not interefere.


Pyrophosphate Assay Kit questions

1. What are the maximum and minimum limits of detection?

The detection limit is ˜0.3 - 1 uM and the Max is 10 uM for ab112155, and 1 mM for ab179836.

2. Are the assays compatible with citrated plasma (plasma collected with citrate)? Read More

We use a buffer containing the following with our samples. Will we be able to use this kit, or will the buffer interfere?

50mM HEPES pH 8, 150mM NaCl, 1mM DDT, 2mM MgCl2

The components of this buffer should not interfere with this assay.

my samples have 3 mM dATP, will this affect the sensitivity of the assay?

Thank you for contacting us. I did contact the lab regarding whether ATP or dATP in samples will compromise the sensitivity of the assay and they did not think it would have much effect. Please let me know if you have any further questions.

For the pyrophosphate assay kit, is it possible to use the kit above room temperature? What is the maximum temperature the kit has been found to work? Thanks.

37C should be no problem. We did not test with a higher temperature, however, 50C might be OK in theory.

I see the probe used in this kit is proprietary in nature but I was wondering if any sturctural information is available for it?

It is a proprietary dye so unfortunately we can't disclose the structure.

preparation of yeast samples? cell lysis?



J'ai reçu la confirmation qu'un tampon de lyse standard pour les levures peut être utilisé avec le kit PPi.
Le laboratoire où a été développé ce kit n'a malheureusement pas de donn&ea...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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