Overlay histogram showing HeLa cells stained with ab184815 (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 (ab184815, 1/500 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.
![ab194815 staining ASH2L in HeLa cells. The cells were fixed with 100% methanol (5 min), 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 ab194815 at a working dilution of 1 in 100 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at a diltuion of 1 in 250 overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.This product also gave a positive signal in 4% formaldehyde (10 min) fixed A549 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_1/10186_ab184815-235840-ap2027610-5ug-ab150120-2ug-HeLam2.jpg)
ab194815 staining ASH2L in HeLa cells. The cells were fixed with 100% methanol (5 min), 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 ab194815 at a working dilution of 1 in 100 (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at a diltuion of 1 in 250 overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.This product also gave a positive signal in 4% formaldehyde (10 min) fixed A549 cells under the same testing conditions.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).