Cyclooxygenase (COX) Activity Assay Kit (Fluorometric) (ab204699)

Overview

  • Product name
    Cyclooxygenase (COX) Activity Assay Kit (Fluorometric)
    See all Cyclooxygenase kits
  • Detection method
    Fluorescent
  • Sample type
    Tissue, Adherent cells, Suspension cells
  • Assay type
    Enzyme activity
  • Assay time
    0h 45m
  • Species reactivity
    Reacts with: Other species, Mammals
  • Product overview

    Cyclooxygenase COX Activity Assay Kit (Fluorometric) (ab204699) provides a simple, sensitive, and high-throughput adaptable method to detect the peroxidase activity of COX in biological samples or purified/crude enzyme preparations. The kit includes COX-1 and COX-2 specific inhibitors to differentiate the activity of COX-1 and COX-2 as well as other peroxidases, which may be present in the sample. Detection limit: 6 μU/mg.


    Cyclooxygenase assay protocol summary:
    - add samples and standards to wells
    - add reaction mix
    - add arachidonic acid/NaOH solution
    - analyze with microplate reader every 15 s for 30 min

  • Notes

    Cyclooxygenase (COX), also known as prostaglandin-endoperoxide synthase (PTGS, EC 1.14.99.1), is an enzyme that is responsible for the formation of important biological mediators called prostanoids, including prostaglandins, prostacyclin and thromboxane. COX is the central enzyme in the biosynthetic pathway to prostanoids from arachidonic acid. There are two known isoenzymes: COX-1 and COX-2. COX-1 is constitutively expressed in many tissues and is the predominant form in gastric mucosa and in kidney. COX-2 is not expressed under normal conditions in most cells, but elevated levels are observed during inflammation. Pharmacological inhibition of COX by nonsteroidal anti-inflammatory drugs (NSAID) can provide relief from the symptoms of inflammation and pain.

  • Platform
    Microplate reader

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 100 tests
    Arachidonic Acid 1 x 50µl
    Celecoxib (COX-2 inhibitor in DMSO) 1 x 100µl
    COX Assay Buffer 1 x 25ml
    COX Cofactor (in DMSO) 1 x 20µl
    COX Probe (in DMSO) 1 x 200µl
    COX-1 Positive Control 1 vial
    NaOH 1 x 500µl
    Resorufin Standard (5 mM, in DMSO) 1 x 50µl
    SC560 (COX-1 inhibitor in DMSO) 1 x 100µl

Images

  • Typical Resorfurin Standard calibration curve.

  • Measurement of COX-1 activity in absence (blue line) and presence of COX-1 inhibitor (SC560, red line) using the COS-1 positive control (PC) provided in the kit.

  • Detection of endogenous COX-1 activity in J774 cell lysate (6 μg) and rat liver homogenate (210 μg).

  • Detection of endogenous COX-2 activity in J774 cell lysate (7 μg) stimulated with or without 100 ng/ml LPS and 100 ng/ml murine IFN-gamma.

Protocols

References

This product has been referenced in:
  • Guenzle J  et al. Cyclooxygenase (COX) Inhibition by Acetyl Salicylic Acid (ASA) Enhances Antitumor Effects of Nitric Oxide in Glioblastoma In Vitro. Mol Neurobiol N/A:N/A (2019). Read more (PubMed: 30715649) »
  • Gilman KE  et al. P2X7 receptor deletion suppresses ?-radiation-induced hyposalivation. Am J Physiol Regul Integr Comp Physiol 316:R687-R696 (2019). Read more (PubMed: 30892913) »
See all 3 Publications for this product

Customer reviews and Q&As

Question
Answer

We do not have any COX activity data from tests of plant extracts. If the plants contain COX that is capable of acting on the arachidonic substrate provided in the kit, then the assay should be able to detect COX activity in those plants, but I do not know whether plants do in fact produce COX or analogs. I found this statement in a review at the following link.


Cyclooxygenases in unicellular organisms, insects, or the plant kingdom have not been identified; however, COX enzymes have recently been determined to be members of a larger fatty acid oxygenase family that includes pathogen-inducible oxygenases (PIOXs).


http://pharmrev.aspetjournals.org/content/56/3/387

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up