Anti-Fatty Acid Synthase 抗体 [EPR7466] - BSA and Azide free (ab221934)

This data was developed using ab221934, the same antibody clone in a different buffer formulation.
Purified ab221934 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 ab221934, the same antibody clone in a different buffer formulation.
Flow Cytometry analysis of A549 (Human lung carcinoma epithelial cell) cells labelling Fatty Acid Synthase with Purified ab221934 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 ab221934, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Fatty Acid Synthase with Purified ab221934 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling Fatty Acid Synthase with Purified ab221934 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 at 1:0 dilution. 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling Fatty Acid Synthase with Purified ab221934 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 at 1:0 dilution. 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling Fatty Acid Synthase with Purified ab221934 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 at 1:0 dilution. PBS instead of the primary antibody was used as the negative control. 

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

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.