![ab93900 staining Hsp90 in HeLa cells. The cells were fixed with 100%methanol (5 min), permeabilised 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 ab193900 at 1/250 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/167 dilution overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.This product gave a positive signal in 100% methanol (5 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_3/5208_ab193900-233551-AP211753801.jpg)
ab93900 staining Hsp90 in HeLa cells. The cells were fixed with 100%methanol (5 min), permeabilised 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 ab193900 at 1/250 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/167 dilution overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.This product gave a positive signal in 100% methanol (5 min) fixed A549 cells under the same testing conditions.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Overlay histogram showing HeLa cells stained with ab193900 (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 (ab193900, 0.2μl/1x106 cells) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Alexa Fluor® 488) (0.1μg/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 20mW Argon ion laser (488nm) and 525/30 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.