Overlay histogram showing HeLa cells stained with ab199253 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab199253, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.
![ab199253 staining p57 Kip2 in SKNSH cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199253 at 1/100 dilution (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/6015_ab199253-239653-ICCAP227543501SKNSH.jpg)
ab199253 staining p57 Kip2 in SKNSH cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199253 at 1/100 dilution (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
![ab199253 staining p57 Kip2 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199253 at 1/100 dilution (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/6016_ab199253-239652-ICCAP227543501.jpg)
ab199253 staining p57 Kip2 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199253 at 1/100 dilution (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).