Anti-c-Myc (phospho S62) antibody [33A12E10] (ab78318)

ab78318 staining c-Myc (phospho S62) in HeLa cells  from Human Cervical Cancer by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with Triton X-100 0.25% in PBS and blocked with 1.5% BSA for 30 minutes at 21°C. Samples were incubated with primary antibody (1/2000 in PBS + 1% BSA) overnight at 4°C. An  Alexa Fluor®488-conjugated Goat anti-mouse IgG polyclonal(1/1000 in 1% BSA) was used as the secondary antibody incudated for 1 hour at room temperature. DAPI was used to stain the nuclear DNA.

Overlay histogram showing HL60 cells stained with ab78318 (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 (ab78318, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HL60 cells fixed in 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.