HTRF-based assay for microsomal prostaglandin E2 synthase-1 activity

Microsomal prostaglandin E-2 synthase-I (mPGES-I) catalyzes the formation of prostaglandin E-2 (PGE(2)) from the endoperoxidase prostaglandin H-2 (PGH(2)). Expression of this enzyme is induced during the inflammatory response,and mouse knockout experiments suggest it may be an attractive target for antoarthritic therapies. Assaying the activity of this enzyme in vitro is challenging because of the enzyme prerparations and for determining the potency of inhibitors compunds. This plate-based competition assay uses homogenous time-resolved fluorescence to measure PGE(2) produced byt he enzyme. The assay is insensitive to DMSO concentration uo to 10% and does not require extensive washes after the initial enzyme reaction is concluded, making it a simple and convenient way to assess mPGES-I inhibition.