Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (AB55178)

ab55178 at 10 ug/ml staining FOXA1 in human HepG2 cells by Immunocytochemistry/ Immunofluorescence.

This image was generated using the ascites version of the product.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (AB55178)

FOXA1 antibody (ab55178) used in immunohistochemistry at 3ug/ml on formalin fixed and paraffin embedded human salivary gland.

This image was generated using the ascites version of the product.

• Flow Cyt

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Flow Cytometry - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (AB55178)

Overlay histogram showing HepG2 cells stained with ab55178 (red line). The cells were fixed with 4 % paraformaldehyde (10 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. The cells were then incubated with the antibody (ab55178, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1μg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed.
This image was generated using the ascites version of the product.

• WB

Lab

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (AB55178)

Lanes 1- 2 : Merged signal (red and green). Green - ab55178 observed at 49 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) observed at 37 kDa.

ab55178 was shown to react with FOXA1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261823 (knockout cell lysate ab256920) was used. Wild-type HeLa and FOXA1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab55178 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

HeLa cells are predicted to express FOXA1, with no expression of FOXA2 or FOXA3.

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (ab55178) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FOXA1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FOXA1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-foxa1-knockout-hela-cell-line-ab261823'>ab261823</a>)

Predicted band size: 49 kDa

Observed band size: 49 kDa

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• WB

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Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (AB55178)

FOXA1 antibody (ab55178) at 1ug/lane + HepG2 cell lysate at 25ug/lane.

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (ab55178)

Predicted band size: 49 kDa

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• WB

Lab

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (AB55178)

Western blot : Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] ab55178 staining at 1/500 dilution, shown in green; Rabbit anti GST (ab111947) loading control staining at 1/200 dilution, shown in magenta. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [1B1] (ab55178) at 1/500 dilution

Lane 1:

FOXA1 Recombinant Protein at 0.1 µg

Lane 2:

FOXA2 Recombinant Protein at 0.1 µg

Lane 3:

FOXA3 Recombinant Protein at 0.1 µg

Secondary

All lanes:

Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 52 kDa,50 kDa,37 kDa

Observed band size: 78 kDa,76 kDa,63 kDa

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