COX2 Mouse Monoclonal Antibody [A3F7]
cat.: EM1902-12
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Rat
Applications: WB, IHC-P, FC
Clonality: Monoclonal
Clone number: A3F7
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 69 kDa
Isotype: IgG1
Immunogen: Synthetic peptide within C-terminal of human COX2.
Positive control: Hela cell lysates ,rat bladder tissue, human lung carcinoma tissue, human womb tissue, A549.
Subcellular location: Microsome membrane, endoplasmic reticulum membrane.
Recommended Dilutions:
  WB
  IHC-P
  FC
1:500
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P35354 Human | 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
EM1902-12_1.jpg Fig1: All lanes: Western blot analysis of COX2 with anti-COX2 antibody [A3F7] (EM1902-12) at 1:1,000 dilution.
Lane 1/2: Wild-type Hela whole cell lysate (20 µg).
Lane 3/4: COX2 knockdown Hela whole cell lysate (20 µg).

EM1902-12 was shown to specifically react with COX2 in wild-type Hela 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 (EM1902-12, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG-HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.
EM1902-12_2.jpg Fig2: Western blot analysis of COX2 on different lysates with Rabbit anti-COX2 antibody (EM1902-12) at 1/2,000 dilution.

Lane 1: HeLa (Human cervical adenocarcinoma cell)

Lysates/proteins at 15 µg/Lane.
Exposure time: 20 seconds; ECL: K1801


Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: EM1902-12, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 69 kDa
Observed band size: 69 kDa
EM1902-12_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat bladder tissue using anti-COX2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-12, 1/100) for 30 minutes 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.
EM1902-12_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-COX2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-12, 1/100) for 30 minutes 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.
EM1902-12_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human womb tissue using anti-COX2 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1902-12, 1/100) for 30 minutes 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.
EM1902-12_6.jpg Fig6: Flow cytometric analysis of COX2 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1902-12, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.