Anti-COX1 / Cyclooxygenase 1 antibody [5F6/F4] (ab695)

Primary antibody incubated for 1 hour at 20°C in 5% milk in TBST. Gel running conditions: Denaturing. Blocked using 5% milk for 16 hours at 4°C. Detection method: Western lightning chemiluminescent reagent.

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Human adjacent breast tissue to tumor or breast tumor tissue labeled by indirect immunofluorescence for COX-1 (red) using ab695, with DAPI as nuclear counterstain (blue).

ab695 at 1/400 staining human anal cancer tissue (Left panel) and mouse kidney tissue (Right panel) sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed before the tissue was incubated with the antibody for 45 minutes. An HRP conjugated goat anti-mouse antibody was used as the secondary.

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Sample: Huh-7 whole cell lysate
Total protein in input: 1x10⁶ cells
Treatment: Cells expressing recombitnant myc-tagged COX1
Immunoprecipitation step: Protein A

ab695 used at 4 µg/mg lysate, inucbated for 4 hours at 4°C.

WB antibody was ab9106 used at a 1/5000 dilution.

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Overlay histogram showing NIH/3T3 (Mouse embryo fibroblast cell line) cells stained with ab695 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. 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 (ab695, 2 µg/1x10⁶ cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2 µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

COX1 protein standard prepared from ram seminal vesicles supplied by Alpha Diagnostic International.

Lane 1 MM.

Lane 2 Sample