COX Fluorescent Activity Assay Kit

Cyclooxygenase (COX, also called Prostaglandin H Synthase or PGHS) is a bifunctional enzyme exhibiting both COX and peroxidase activities. The COX component converts arachidonic acid to a hydroperoxy endoperoxide (PGG₂), and the peroxidase component reduces the endoperoxide to the corresponding alcohol (PGH₂), the precursor of PGs, thromboxanes, and prostacyclins.^1,2 It is now well established that there are two distinct isoforms of COX, namely COX-1 and COX-2. Cayman’s COX Fluorescent Activity Assay provides a convenient fluorescence-based method for detecting COX-1 or COX-2 activity in both crude (cell lysates/tissue homogenates) and purified enzyme preparations. The assay utilizes the peroxidase component of COXs. In this assay, the reaction between PGG₂ and ADHP (10-acetyl-3,7-dihydroxyphenoxazine) produces the highly fluorescent compound resorufin that can be analyzed using an excitation wavelength of 530-540 nm and an emission wavelength of 585-595 nm. The kit includes isozyme-specific inhibitors for distinguishing COX-2 activity from COX-1 activity.

Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.

WARNING This product is not for human or veterinary use.

1. Nugteren, D.H., and Hazelhof, E. Isolation and properties of intermediates in prostaglandin biosynthesis. Biochim. Biophys. Acta 326(3), 448-461 (1973).

2. Hamberg, M., and Samuelsson, B. Detection and isolation of an endoperoxide intermediate in prostaglandin biosynthesis. Proc. Natl. Acad. Sci. USA 70(3), 899-903 (1973).

Nam, G.S., Kim, S., Kwon, Y.-S., et al. A new function for MAP4K4 inhibitors during platelet aggregation and platelet-mediated clot retraction. Biochem. Pharmacol. 188, 114519 (2021).

Kugathas, S., Audouze, K., Ermler, S., et al. Effects of common pesticides on prostaglandin D₂ (PGD₂) Inhibition in SC5 mouse sertoli cells, evidence of binding at the COX-2 active site, and implications for endocrine disruption. Environ. Health Perspect. 124(4), 452-459 (2016).

Pang, L., Cai, Y., Tang, E.H., et al. Cox-2 inhibition protects against hypoxia/reoxygenation-induced cardiomyocyte apoptosis via Akt-dependent enhancement of iNOS expression. Oxid. Med. Cell. Longev. 3453059 (2016).