Anti-Nucleolin antibody (ab22758)

ICC/IF image of ab22758 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab22758, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iT^TM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

ab22758 staining nucleolin in HeLa cells treated with Triptolide from Tripterygium wilfordii (ab120720), by ICC/IF. Changes in nuclear localization of nucleolin (from nuclelous to whole nucleous) correlates with increased concentration of Triptolide from Tripterygium wilfordii, as described in literature.
The cells were incubated at 37°C for 1h in media containing different concentrations of ab120720 (Triptolide from Tripterygium wilfordii ) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab22758 (1 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.