Pyrophosphate Assay Kit (Fluorometric) (ab112155)

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).

  • Tested applications
    Suitable for: Functional Studiesmore details

Properties

    Applications

    Our Abpromise guarantee covers the use of ab112155 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Abreviews Notes
    Functional Studies Use at an assay dependent concentration.

    Pyrophosphate Assay Kit (Fluorometric) images

    • 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.

    • 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 for Pyrophosphate Assay Kit (Fluorometric) (ab112155)

    ab112155 has not yet been referenced specifically in any publications.

    Product Wall

    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 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).
    Username

    Abcam user community

    Verified customer

    Submitted Jan 19 2017

    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.


    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

    The components of this buffer should not interfere with this 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.

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

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



    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...

    Read More

    1-10 of 30 Abreviews or Q&A

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

    Sign up