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

ab192467 staining Cytokeratin 8 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

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)

Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab192467 (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 (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.