COX2/Cyclooxygenase 2 Recombinant Rabbit Monoclonal Antibody [SC56-06]
cat.: ET1610-23
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SC56-06
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 69 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human COX2 aa 100-140.
Positive control: NIH/3T3 cell lysate, C2C12 cell lysate, L6 cell lysate, Mouse bladder tissue lysate, Mouse small intestine tissue lysate, C2C12, A549 cell lysates, A549, rat bladder tissue, human colon tissue, mouse kidney tissue, mouse uterus tissue.
Subcellular location: Microsome membrane, Endoplasmic reticulum membrane, Nucleus inner membrane, Nucleus outer membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:500-1:1,000
1:50-1:200
1:50-1:200
1:100-1:5,000
Uniprot #: SwissProt: P35354 Human | Q05769 Mouse | P35355 Rat
Alternative names: COX 2 COX-2 COX2 Cyclooxygenase 2 Cyclooxygenase 2b Cyclooxygenase Cyclooxygenase-2 Cyclooxygenase2 EC 1.14.99.1 fj02a10 Glucocorticoid-regulated inflammatory cyclooxygenase Glucocorticoid-regulated inflammatory Prostaglandin G/H synthase GRIPGHS hCox 2 Macrophage activation-associated marker protein P71/73 OTTHUMP00000033524 PES-2 PGG/HS PGH synthase 2 PGH2_HUMAN PGHS 2 PGHS-2 PGHS2 PHS 2 PHS II PHS2 Prostaglandin endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) Prostaglandin endoperoxide synthase 2 Prostaglandin G/H synthase 2 Prostaglandin G/H synthase 2 precursor Prostaglandin G/H synthase and cyclooxygenase Prostaglandin G/H synthase Prostaglandin H2 synthase 2 prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) Prostaglandin-endoperoxide synthase 2 PTGS2 ptgs2a TIS10 TIS10 protein unp1239 wu:fj02a10
Images
ET1610-23_1.jpg Fig1: Western blot analysis of COX2/Cyclooxygenase 2 on different lysates with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: C2C12 cell lysate (20 µg/Lane)
Lane 3: L6 cell lysate (20 µg/Lane)
Lane 4: Mouse bladder tissue lysate (40 µg/Lane)
Lane 5: Mouse small intestine tissue lysate (40 µg/Lane)

Predicted band size: 69 kDa
Observed band size: 69 kDa

Exposure time: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-23) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1610-23_2.jpg Fig2: Immunocytochemistry analysis of C2C12 cells labeling COX2/Cyclooxygenase 2 with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1610-23_3.jpg Fig3: All lanes: Western blot analysis of COX2 with anti-COX2 antibody [SC56-06] (ET1610-23) at 1:1,000 dilution.
Lane 1/2: Wild-type A431 whole cell lysate (20 µg).
Lane 3/4: COX2 fragment knockdown A431 whole cell lysate (20 µg).

ET1610-23 was shown to specifically react with COX2 in wild-type A431 cells. Weakened bands were observed when COX2 knockdown samples were tested. Wild-type and COX2 knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-23, 1/1,000) and Loading control antibody (Rabbit anti-Vinculin, ET1705-94, 1/5,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
ET1610-23_4.jpg Fig4: Western blot analysis of COX2/Cyclooxygenase 2 on A549 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-23, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
ET1610-23_5.jpg Fig5: Immunocytochemistry analysis of A549 cells labeling COX2/Cyclooxygenase 2 with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
ET1610-23_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded rat bladder tissue with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-23) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-23_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-23) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-23_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-23) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-23_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded mouse uterus tissue with Rabbit anti-COX2/Cyclooxygenase 2 antibody (ET1610-23) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-23) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.