Overlay histogram showing intracellular staining of HeLa cells labeled with ab52459 (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 (ab52459, 1µg/1x10⁶ cells) for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 (FITC) (2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

ICC/IF image of ab52459 stained HCT116 cells. The cells were 4% formaldehyde fixed (10 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 (ab52459, 5µg/ml) overnight at +4°C. 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.

Flow cytometry (Intracellular) analysis HeLa (human cervix carcinoma cell line) cells labeling Cytokeratin 18 with ab52459 (red). Overlay with Isotype mouse IgG1 control FITC antibody (blue).