Recombinant Anti-COX1 / Cyclooxygenase 1 antibody [EPR5866] (ab109025)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling COX1 / Cyclooxygenase 1 with Purified ab109025 at 1:150 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution.

PBS instead of the primary antibody was used as the negative control.

Blocking and diluting buffer: 5% NFDM/TBST.

Blocking and diluting buffer: 5% NFDM/TBST.

ab109025 (purified) at 1:20 dilution (0.8μg) immunoprecipitating COX1 / Cyclooxygenase 1 in C2C12 whole cell lysate.

Lane 1 (input): C2C12 (Mouse myoblasts myoblast) whole cell lysate,10μg
Lane 2 (+): ab109025 & C2C12 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109025 in C2C12 whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.

Blocking and diluting buffer: 5% NFDM/TBST.

Intracellular Flow Cytometry analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling COX1 / Cyclooxygenase 1 with purified ab109025 at 1/100 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling COX1 / Cyclooxygenase 1 with Purified ab109025 at 1:150 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution.

PBS instead of the primary antibody was used as the negative control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum tissue sections labeling COX1 / Cyclooxygenase 1 with Purified ab109025 at 1:150 dilution. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution.

PBS instead of the primary antibody was used as the negative control.

Immunocytochemistry/Immunofluorescence analysis of Neuro-2a (mouse neuroblastoma) labelling COX1 with purified ab109025 at 1/50. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

Control: PBS only.

Unpurified ab109025 at 1/250 dilution staining COX1 / Cyclooxygenase 1 in human skin by immunohistochemistry, paraffin-embedded tissue.

Unpurified ab109025 at 1/100 dilution staining COX1 / Cyclooxygenase 1 in HeLa cells by Immunofluorescence.

Overlay histogram showing NIH/3T3 cells stained with unpurified ab109025 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109025, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.