Overlay histogram showing HeLa cells stained with ab198981 (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 (ab198981, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 488 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 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
![ab198981 staining SUN2 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 ab198981 at 1/100 dilution (shown in green) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed, at 1µg/ml (shown in red). 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_5/6849_ab198981-239499-ICCAP226937001SKNSH.jpg)
ab198981 staining SUN2 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 ab198981 at 1/100 dilution (shown in green) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed, at 1µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
![ab198981 staining SUN2 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 ab198981 at 1/50 dilution (shown in green) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed, at 1µg/ml (shown in red). 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_5/6850_ab198981-239498-ICCAP226937001.jpg)
ab198981 staining SUN2 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 ab198981 at 1/50 dilution (shown in green) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml. This was followed by an incubation at room temperature for 1h with ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed, at 1µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).