Cox 2 activity assay kit

1α,25(OH)₂D₃, and the antibiotic G418 were from Sigma-Aldrich (St. Louis, MO, USA). Puromycin was provided by Invivogen (San Diego, CA, USA). The antibodies used were rat monoclonal anti-VDR (Affinity Bioreagents, Golden, CO, USA); mouse monoclonal anti-COX-2, anti-mouse and anti-rat horseradish peroxidase–conjugated secondary antibody (Santa Cruz, CA, USA). Roche Applied Science (Indianapolis, IN, USA) provided high Pure RNA Isolation Kit. Immobilon P (polyvinylidene difluoride; PVDF) membranes were from Thermo Scientific (Rockford, IL, USA); PCR primers for mouse Gapdh, Cox-2, EP1, EP2, EP3 and EP4 were synthetized by Invitrogen (Thermo Scientific Inc., Rockford, IL, USA). Celecoxib (Santa Cruz, CA, USA). COX-2 Activity Assay kit (Cayman N° 760151) was from Cayman Chemical Company (Michigan, USA).

COX activity was measured by COX-2 Activity Assay kit (Cayman N° 760151) following the recommendations provided by the manufacturer. In time response-experiments protein content from whole cell lysates was determined by the Bradford procedure to standardize the signal detected [22 (link)]. The product of the reaction was colorimetrically measured at 590 nm.