![Ab195303 staining GM130 in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab195303 at 5μg/ml (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1/100 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Mouse secondary (ab150117) at 2μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.This product gave a positive signal in 4% formaldehyde (10min) fixed HeLa cells under the same testing conditions.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/26000_ab195303-234733-ICCAP21503531.jpg)
Ab195303 staining GM130 in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab195303 at 5μg/ml (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1/100 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with an AlexaFluor®488 Goat anti-Mouse secondary (ab150117) at 2μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.This product gave a positive signal in 4% formaldehyde (10min) fixed HeLa cells under the same testing conditions.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Overlay histogram showing HeLa cells stained with ab195303 (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 (ab195303, 0.1μg/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Alexa® 647) (0.2μl/1x106 cells) for 30 min at 22ºC. Unlabelled sample (blue line) was also used as a control.Acquisition of >5,000 events were collected using a 25mW red solid state diode laser (635nm) and 675/30 bandpass filter.This antibody gave a positive signal in HeLa fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.