Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Fatty Acid Synthase knockout HAP1 cell lysate (20 µg)
Lane 3: A549 cell lysate (20 µg)
Lane 4: Hu liver tissue lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab128870 observed at 250 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab128870 was shown to react with Fatty Acid Synthase in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when Fatty Acid Synthase knockout samples were examined. Wild-type and Fatty Acid Synthase knockout samples were subjected to SDS-PAGE. ab128870 and ab18058 (loading control to Vinculin) were both diluted at 1/10,000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

Overlay histogram showing A549 cells stained with ab128870 (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 (ab128870, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. 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 A549 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

All lanes : Anti-Fatty Acid Synthase antibody [EPR7466] (ab128870) at 1/10000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : 293T cell lysate
Lane 3 : A549 cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

Developed using the ECL technique.

Predicted band size: 273 kDa

ab128870, at a 1/100 dilution, staining Fatty Acid Synthase in paraffin embedded Human liver tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab128870, at a 1/250 dilution, staining Fatty Acid Synthase in A549 cells by Immunofluorescence.

Equilibrium disassociation constant (K_D)
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