ab17139 staining human mammacarcinoma by IHC-P.
Overlay histogram showing MCF7 cells stained with ab17139 (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. The cells were then incubated with the antibody (ab17139, 1/100 dilution) 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. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
ab17139 staining Cytokeratin 8+18 in MCF7 cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in paraformaldehyde and permeabilized in 0.1% Triton X-100 prior to blocking in 1% BSA for 1 hour at room temperature. The primary antibody was diluted 1/1000 and incubated with the sample for 1 hour at room temperature. The secondary antibody was Alexa Fluor 488®-conjugated Goat anti-Mouse, diluted 1/1000.See Abreview