Lane 1: MM standardLane 2: Cox-1 (protein standard prepared from ram seminal vesicles, supplied by Alpha Diagnostic International)Antibody 5F6/F4: 1 µg/mlLane 1: MM standardLane 2: Cox-1 (protein standard prepared from ram seminal vesicles, supplied by Alpha Diagnostic International)Antibody 5F6/F4: 1 µg/ml
ab695 at 1/400 staining human anal cancer tissue 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.See Abreview
Anti-COX1 / Cyclooxygenase 1 antibody [5F6/F4] (ab695) at 1 µg/ml + whole cell lysate prepared from human huh-7 cell line at 15 µgSecondarySheep anti-mouse IgG conjugated to HRP at 1/1000 dilutionPerformed under reducing conditions.Observed band size : 68 kDa (why is the actual band size different from the predicted?)Additional bands at : 38 kDa. We are unsure as to the identity of these extra bands.Exposure time : 4 minutesThis image is courtesy of an anonymous abreview.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.See Abreview
Overlay histogram showing NIH3T3 cells stained with ab695 (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 (ab695, 2µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
ab695 staining COX1 / Cyclooxygenase 1 in the Mouse embryonic fibroblasts (MEF)cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Triton X-100 1% in PBS and blocked with 5% serum for 60 minutes at 21°C. Samples were incubated with primary antibody (1/100 in 5% Goat serum) for 2 hours. An Alexa Fluor®546-conjugated Goat anti-mouse IgG polyclonal(1/400) was used as the secondary antibody.See Abreview
ab695 staining COX1 / Cyclooxygenase 1 in Mouse heart tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 60 minutes at 25°C. Samples were incubated with primary antibody (1/200 in 5% Goat serum) for 2 hours. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.See Abreview