Key features and details
- Assay type: Enzyme activity
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 45 min
- Sample type: Adherent cells, Suspension cells, Tissue
Product nameCyclooxygenase (COX) Activity Assay Kit (Fluorometric)
See all Cyclooxygenase kits
Sample typeTissue, Adherent cells, Suspension cells
Assay typeEnzyme activity
Assay time0h 45m
Species reactivityReacts with: Mammals, Other species
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
Cyclooxygenase (COX), also known as prostaglandin-endoperoxide synthase (PTGS, EC 18.104.22.168), 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.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 100 tests Arachidonic Acid 1 vial 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
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.
ab204699 has been referenced in 6 publications.
- Mauri E et al. Ester coupling of ibuprofen in hydrogel matrix: A facile one-step strategy for controlled anti-inflammatory drug release. Eur J Pharm Biopharm 146:143-149 (2020). PubMed: 31726217
- Wang J et al. Betalain exerts a protective effect against glaucoma is majorly through the association of inflammatory cytokines. AMB Express 10:125 (2020). PubMed: 32666339
- Montaser A et al. Astrocyte-Targeted Transporter-Utilizing Derivatives of Ferulic Acid Can Have Multifunctional Effects Ameliorating Inflammation and Oxidative Stress in the Brain. Oxid Med Cell Longev 2019:3528148 (2019). PubMed: 31814871
- 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). PubMed: 30715649
- Gilman KE et al. P2X7 receptor deletion suppresses ?-radiation-induced hyposalivation. Am J Physiol Regul Integr Comp Physiol 316:R687-R696 (2019). PubMed: 30892913
- Destefanis F et al. Investigating the Connection Between Endogenous Heme Accumulation and COX2 Activity in Cancer Cells. Front Oncol 9:162 (2019). PubMed: 30941311