Anti-Fatty Acid Synthase antibody [EPR7465]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Fatty Acid Synthase antibody [EPR7465] (AB128856)

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Fatty Acid Synthase with Purified ab128856 at 1 : 50 dilution (4.5 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Fatty Acid Synthase antibody [EPR7465] (AB128856)

Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labelling Fatty Acid Synthase with Purified ab128856 at 1 : 50 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1 : 2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• WB

Unknown

Western blot - Anti-Fatty Acid Synthase antibody [EPR7465] (AB128856)

All lanes:

Western blot - Anti-Fatty Acid Synthase antibody [EPR7465] (ab128856) at 1/5000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg

Lane 5:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

Mouse brain lysate at 20 µg

Lane 7:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 8:

Rat brain lysate at 20 µg

Lane 9:

L6 (Rat skeletal muscle myoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 273 kDa

Observed band size: 237 kDa,273 kDa

false

• WB

Lab

Western blot - Anti-Fatty Acid Synthase antibody [EPR7465] (AB128856)

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 - ab128856 observed at 250 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab128856 was shown to specifically react with Fatty Acid Synthase in wild-type HAP1 cells. 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. ab128856 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10,000 respectively 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.

All lanes:

Western blot - Anti-Fatty Acid Synthase antibody [EPR7465] (ab128856)

Predicted band size: 273 kDa

false