| Assay Method Information | |
| | Cyclooxygenase Assay |
| Description: | 100 mM Tris HCl buffer, pH 8.0 containing 1 µM heme and COX-1 (ovine) or COX-2(human recombinant), which was preincubated for 10 min in a water bath at 37 °C was used for reaction mixture preparation. Addition of 10 µL arachidonic acid to the reaction mixture (final concentration 100 µM) was used to initiate reaction. After 2 min reaction was stopped by addition of 1 M HCl. PGE2 in the reaction mixture was determined using ELISA method. 100 mM potassium phosphate buffer (pH 7.4) was used to dilute the DMSO stock solutions of the test compounds to the desired concentration. This reaction mixture was transferred to a 96-well platethat was precoated with a mouse anti-rabbit IgG. Tracer prostaglandin acetylcholine esterase and primary antibody (mouse anti PGE2) were also added to the plates followed by overnight incubation at room temperature. After overnightincubation, reaction mixtures were pipetted out and the wells were washed with 10 mM potassium phosphate buffer containing 0.05% Twe |
| Affinity data for this assay | |
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