Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Fatty Acid Synthase with Purified ab128870 at 1 : 50 dilution (10 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1 : 200 (2.5 μg/ml). 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.

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labelling Fatty Acid Synthase with Purified ab128870 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).

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling Fatty Acid Synthase with Purified ab128870 at 1 : 450 dilution (1.09 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128870).

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and FASN knockout HAP1 stained with ab128870 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab128870) (1x 10⁶ in 100μl at 0.008 μg/ml (1/69250 dilution)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in HAP1 WT cells (black line) and HAP1-FASN KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This antibody gave a positive signal in wild-type HAP1 fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• IP

Lab

Immunoprecipitation - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

Purified ab128870 at 1 : 30 dilution (2μg) immunoprecipitating Fatty Acid Synthase in HEK-293 whole cell lysate.
Lane 1 (input) : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10 μg
Lane 2 (+) : ab221934 + HEK-293 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab128870 in HEK-293 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP)(ab131366) (1 : 10,000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : kDa

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

All lanes:

Immunoprecipitation - Anti-Fatty Acid Synthase antibody [EPR7466] (<a href='/en-us/products/primary-antibodies/fatty-acid-synthase-antibody-epr7466-ab128870'>ab128870</a>)

Predicted band size: 273 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling with ab128870 at 1/50 dilution, followed by ab150081 antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cells is observed. ab195889 was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Fatty Acid Synthase with Purified ab128870 at 1 : 450 dilution (1.09 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Fatty Acid Synthase with Purified ab128870 at 1 : 450 dilution (1.09 μg/ml). Heat mediated antigen retrieval was performed using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

• WB

Supplier Data

Western blot - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

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.

This image was produced using ab128870, the same clone but in a different formulation.

All lanes:

Western blot - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (ab221934)

Predicted band size: 273 kDa

false

• WB

Unknown

Western blot - Anti-Fatty Acid Synthase antibody [EPR7466] - BSA and Azide free (AB221934)

This data was developed using ab128870, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Fatty Acid Synthase antibody [EPR7466] (<a href='/en-us/products/primary-antibodies/fatty-acid-synthase-antibody-epr7466-ab128870'>ab128870</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

Mouse liver lysate at 20 µg

Lane 4:

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

Lane 5:

Mouse brain lysate at 20 µg

Lane 6:

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

Lane 7:

Rat brain lysate at 20 µg

Lane 8:

Rat liver lysate at 20 µg

Lane 9:

HEK-293 (Human embryonic kidney epithelial cell) 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

false