Cox 2 antibody

Tumor homogenates were harvested from mice treated as indicated, and mechanically disaggregated and digested with triple enzyme mixture (Collagenase type IV, DNase type IV, and Hyaluronidase type V (Sigma-Aldrich, St Louis, MO)). Serum was collected through sub-mandibular bleed. In vitro cells infected with virus at MOI=1 for 24 hr or pretreated for 24 hr with 20 nm Celecoxib (BioVision, San Francisco, CA) were pretreated with 20 μm arachidonic acid (Sigma-Aldrich, St. Louis, MO) 4 hr prior to harvest. All experiments were performed in triplicate. Mouse SDF-1(CXCL12), RANTES (CCL5) and I-TAC(CXCL11) ELISA were performed according to the manufacturer’s protocol (Abcam, Cambridge, MA) and PGE2 ELISA were performed according to the manufacturer’s protocol (Cayman, Ann Arbor, Michigan) and optical density was detected with the 3,3,5,5-tetramethylbenzidine (TMB) peroxidase substrate kit (Vector Labs, Burlingame, CA).
For western blot assay, in vitro lysed cell protein was prepared. Mouse HPGD antibody (Abcam,Cambridge, MA), COX2 antibody (BD, Franklin Lakes, New Jersey) and beta-actin antibody (Sigma-Aldrich, St. Louis, MO) were used for Western Blot according to the manufacturer’s protocol.