ab76548 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab76548 at 10µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-mouse DyLight® 488 (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in methanol fixed (100%, 5min) Hek293, HepG2, and MCF-7 cells at 10ug/ml.
![Anti-Actin antibody [ACTN05; same as C4] (ab76548) at 0.5 µg/ml + Human cells 8](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/2154_panactin.jpg)
Anti-Actin antibody [ACTN05; same as C4] (ab76548) at 0.5 µg/ml + Human cells 8
Actin was immmunoprecipitated from human cells 8 using ab76548, followed by Western blotting using ab76548 at 0.5 µg/ml.
![Overlay histogram showing HeLa cells stained with ab76548 (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 (ab76548, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/2156_Actin-Primary-antibodies-ab76548-1.jpg)
Overlay histogram showing HeLa cells stained with ab76548 (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 (ab76548, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.