Recombinant Alexa Fluor® 488 Anti-Cytokeratin 8 antibody [EP1628Y] (ab192467)

Immunofluorescence analysis of mouse myoepithelial cells labelling KRT8 (right) with ab192467 at 1/100 dilution. The tissue was fixed in 10% neutral buffered formalin overnight. Paraffin embedding and sectioning were performed by the Rodent Histopathology Core at Harvard Medical School. Scale bar, 10 μm.

Immunofluorescence analysis of mouse postate tumor samples labelling cytokeratin 8 (green) with ab192467 at 1/100 dilution. SYP was also stained using ab206870 (red). Cells were fixed with formalin and embedded in paraffin. Sections were blocked with PBS-Tween (0.1%) containing 5% of BSA. Primary conjugated antibodies were simultaneously incubated overnight at 4°C. Nuclear DNA was labelled with DAPI (blue). Scale bar = 100 µm.

Immunofluorescence staining of cytokeratin 8 in HeLa cells using ab192467. The cells were fixed with 4% formaldehyde (10 min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton X-100 for 1hr. The cells were then incubated with ab192647 at a working dilution of 1 in 100 (shown in green) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1hr with AlexaFluor^® 594 Goat anti-mouse IgG (H&L - preadsorbed) (ab150120) at 2 μg/ml (shown in pseudo-color red).

Nuclear DNA was labeled in blue with DAPI.

This product gave a positive signal in 100% methanol (5 min) fixed HeLa cells under the same testing conditions.

Image was taken with a Confocal microscope (Leica-microsystems, TCS SP8)

Flow cytometry analysis of HeLa cells stained with ab192467. 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 (ab192467, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit monoclonal IgG [EPR25A] Alexa Fluor^® 488 (ab199091) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

Immunofluorescence staining in human mammary organoids of cytokeratin 8 using ab192467 (green), cytokeratin 14 using ab206100 (red), and actin (white). Cells were fixed with 4% paraformaldehyde for 20 minutes at room temperature and permeabilized with 0.5% Triton X-100 for 10 minutes at 4 °C. Primary antibodies incubated overnight at 4 °C. Nuclear DNA was labelled with DAPI (blue). Scale bar = 100 µm. Organoids were imaged by confocal microscopy.