ICC/IF image of ab8068 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8068, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Ab8068 staining pan Keratin in Human glioblastoma cell line by ICC/IF (Immunocytochemistry/Immunofluorescence). The cells were fixed with paraformaldehyde; permeabilized with 0.1% Triton X 100 in PBS and blocked with 0.5% BSA for 30 minutes at room temperature. Samples were incubated with primary antibody (1/50 dilution in 0.5% BSA in PBS) for 16hours at 4°C. An Alexa Fluor^® 488 anti-mouse was used as a secondary antibody at 1/400 dilution.

See Abreview

Anti-pan Keratin antibody [80] (ab8068) at 1/250 dilution + A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 65 kDa
Observed band size: 65 kDa
Additional bands at: 49 kDa, 58 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 30 seconds

Overlay histogram showing A431 cells stained with ab8068 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab8068, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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.