Overview

  • Product name

    Cyclooxygenase (COX) Activity Assay Kit (Luminometric)
    See all Cyclooxygenase kits
  • Detection method

    Luminescent
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    COX 16 30U/ml 7%
    COX 16 7.5U/ml 3.1%
  • Assay type

    Enzyme activity
  • Sensitivity

    0.147 U/ml
  • Range

    7.5 U/ml - 60 U/ml
  • Product overview

    The Cyclooxygenase (COX) Activity Assay Kit (Luminometric) (ab139432) is a non-radioactive COX activity assay. This kit allows user to easily evaluate COX I and COX II inhibitors in a convenient 96-well plate based assay with < 7% intra-assay variation.

    COX Activity Assay Kit (Luminometric) uses a specific chemiluminescent substrate to detect the peroxidative activity of COX enzymes. After inhibition by NSAIDs, the direct residual activity of COX is measured by addition of a proprietary luminescent substrate and arachidonic acid. Light emission starts immediately and is directly proportional to the COX activity in the sample. The chemiluminescent signal is measured over 5 seconds.

  • Notes

    COX (cyclooxygenase, also known as Prostaglandin G/H synthase) is a membrane-bound enzyme responsible for the oxidation of arachidonic acid to Prostaglandin G2 (PGG2) and the subsequent reduction of PGG2 to PGH2. These reactions are the first steps in the formation of a variety of prostanoids.

    COX has been shown to be expressed in at least two different isoforms: a constitutively expressed form, COX1, and an inducible form, COX2. COX1 is thought to regulate a number of "housekeeping" functions such as vascular hemostasis and renal blood flow. Inflammation mediators such as growth factors, cytokines and endotoxin induce COX2 expression in a number of cellular systems.

  • Platform

    Microplate reader

Properties

Images

  • Typical standard curve for COX 2 activity (in RLU) following the procedure described in the protocol for ab139432.
  • Typical standard curve for COX 1 activity (in RLU) following the procedure described in the protocol for ab139432.

Protocols

References

This product has been referenced in:

  • Estrada-Figueroa LA  et al. Leishmania mexicana gp63 is the enzyme responsible for cyclooxygenase (COX) activity in this parasitic protozoa. Biochimie 151:73-84 (2018). Read more (PubMed: 29864507) »
See 1 Publication for this product

Customer reviews and Q&As

Answer

This kit hasn't been specifically validated for use with cell lysates, however we do expect it to work if there is sufficient COX activity in the lysate. For every unique sample, the sample handling, treatment, dilution/concentration of sample required, etc will need to be optimized. The protease inhibitors and detergents that we have tested minimally affect the assay, while tissue culture media, PBS, and DMF show more effect. Protease inhibitors (Peafabloc, pepstatin, leupeptin, E-64, bestatin, and aprotinin) at 0.1% in solution gave a 3.3% change in luminescent signal. Tween-20 at 0.1% showed a 5% change in signal. 100% PBS and culture media showed 15.9 and 14.8% signal change respectively. 1% DMF showed a 12.7% signal change. Ethanol, methanol, and 10% DMSO are the solvents that showed the lowest affect on the signal.

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