COX-2 activity was determined using a COX-2 activity assay kit (Cayman Chemical, Ann Arbor, MI) according to the manufacturer’s protocol, as previously described (Sribnick et al. 2010 (link)). Briefly, cells (1×106) were homogenized in cold buffer 0.1 M Tris-HCl, pH 7.8 containing 1 mM EDTA, mixed with assay buffer, heme, and COX-1 inhibitor. Colorimetric substrate and arachidonic acid were added sequentially and incubated at 37°C for 10 min, and the absorbance was read at 590 nm.
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Determination of COX-2 Activity Assay
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Corresponding Organization : University of Alabama at Birmingham
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Variable analysis
independent variables
- COX-2 activity
- Absorbance at 590 nm
- Cell count (1×10^6 cells)
- Homogenization buffer (0.1 M Tris-HCl, pH 7.8 containing 1 mM EDTA)
- Assay buffer
- COX-1 inhibitor
- Colorimetric substrate
- Arachidonic acid
- Incubation time (10 min)
- Incubation temperature (37°C)
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