All lanes : Anti-FOXO3A antibody [EPR1950] (ab109629)

Lane 1 : Wild-type HEK 293 whole cell lysate
Lane 2 : FOXO3 knockout HEK 293 whole cell lysate
Lane 3 : Hela whole cell lysate
Lane 4 : Jurkat whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 71 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab109629 observed at 71 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109629 was shown to recognize FOXO3A in wild-type HEK 293 cells as signal was lost at the expected MW in FOXO3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FOXO3 knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab109629 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-FOXO3A antibody [EPR1950] (ab109629) at 1/1000 dilution

Lane 1 : MCF7 cell lysate
Lane 2 : SH-SY5Y cell lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 71 kDa

Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling FOXO3A with unpurified ab109629 at 1/150 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.