Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution. Heat mediated antigen retrieval was performed using citrate buffer pH 6.0 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunocytochemistry/ Immunofluorescence analysis of PC-3 (Human prostate adenocarcinoma epithelial cell) cells labeling FOXA1 with Purified ab170933 at 1 : 100 dilution (11 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 nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunofluorescence analysis of HepG2 cells labeling FOXA1 with unpurified ab170933 at 1/100 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling FOXA1 with Purified ab170933 at 1 : 1000 dilution (1.13 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstain.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Intracellular Flow Cytometry analysis of PC-3 (human prostate adenocarcinoma epithelial cell) cells labeling FOXA1 with purified ab170933 at 1/50 dilution (1 ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) (1/5000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution. Heat mediated antigen retrieval was performed using citrate buffer pH 6.0 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse liver tissue sections labeling FOXA1 with Purified ab170933 at 1 : 1000 dilution (1.13 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat pancreas tissue sections labeling FOXA1 with Purified ab170933 at 1 : 1000 dilution (1.13 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0) ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody.Negative control : PBS instead of the primary antibody.Hematoxylinwas used as a counterstain

• WB

Unknown

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (ab170933) at 1.1 µg/mL

Lane 1:

SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

Mouse lung lysates at 15 µg

Lane 3:

Rat lung lysates at 15 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: 49 kDa

Observed band size: 49 kDa

false

• WB

Lab

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Lanes 1 - 4 : Merged signal (red and green). Green - ab170933 observed at 52 kDa. Red - loading control, ab9484, observed at 37 kDa.

Unpurified ab170933 was shown to specifically react with FOXA1 in wild-type HAP1 cells as signal was lost in FOXA1 knockout cells. Wild-type and FOXA1 knockout samples were subjected to SDS-PAGE. ab170933 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

HAP1 cells are predicted to express FOXA1, not FOXA2 and FOXA3

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (ab170933) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

FOXA1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

MCF7 whole cell lysate at 20 µg

Lane 4:

HepG2 whole cell lysate at 20 µg

Predicted band size: 49 kDa

false

• WB

Lab

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

Lanes 1- 2 : Merged signal (red and green). Green - ab170933 observed at 52 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab170933 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. ab170933 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

Hela cells are predicted to express FOXA1, not FOXA2 and FOXA3

All lanes:

Western blot - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (ab170933) at 1/1000 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: 52 kDa

false

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab170933 [EPR10881]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab170933 [EPR10881]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

• ChIC/CUT&RUN-seq

Lab

ChIC/CUT&RUN sequencing - Anti-FOXA1 + FOXA2 + FOXA3 antibody [EPR10881] (AB170933)

ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (human hepatocellular carcinoma epithelial cell) cells and 5 µg of ab170933 [EPR10881]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.