COX2 Mouse Monoclonal Antibody [A3F7]

☑ Knockdown (KD)

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.

Western blot analysis of COX2 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 (EM1902-12, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

Western blot analysis of COX2 on rat spinal cord tissue lysates with Mouse anti-COX2 antibody (EM1902-12) at 1/500 dilution.

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 2 minutes;

8% 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 (EM1902-12) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

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.

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.

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.

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).