Cox 1 ovine and cox 2 human inhibitor screening assay kit no 701050

The ability of OLE, EVOO, and EVOO/OLE extracts to inhibit COX-1 and COX-2 was evaluated using a COX-1 (ovine) and COX-2 (human)-inhibitor screening assay (kit No. 701050 from Cayman Chemical Co. Michigan, USA), following the manufacturer’s protocols. An initial COX-1/COX-2 inhibitory evaluation test was performed at 225 µg/mL for all the extracts (OLE, EVO, and EVO/OLE 8%). Then, increasing concentrations of each sample (45–225 µg/mL) have been tested. Arachidonic acid at 1.1 mM was the substrate, and ibuprofene was used as a control. The peroxidase activity was examined colorimetrically at 590 nm after an incubation of 120 min at room temperature using a SPECTROstarNano (200–1000 nm) UV/Vis spectrophotometer (BMG Labtec, Germany). All tests were performed three times. The percent (%) inhibition of COX-1 and COX-2 is derived from the following formula:


Results were expressed as % of inhibition or inhibitory concentration at 50% (IC₅₀) calculated by least-squares regression analysis of inhibition versus concentration, as already reported [8 (link),42 (link),43 (link)]. All experiments were performed in triplicate.

All compounds described here were tested for their ability to inhibit COX-1 and COX-2 using a COX-1(ovine) and COX-2 (human)-inhibitor screening assay kit No. 701050 from Cayman Chemical Co., according to the manufacturer’s instructions. The compounds (1–6) at 70 μM and selective COX-1 inhibitor, SC-560 (3.3 μM) and selective COX-2 inhibitor, DuP-697 (3 μM) dissolved in DMSO or ethanol were assayed.