Validated using a knockout cell line
Recombinant
RabMAb

Anti-FOXA1 antibody [EPR10881] (ab170933)

Overview

  • Product name
    Anti-FOXA1 antibody [EPR10881]
    See all FOXA1 primary antibodies
  • Description
    Rabbit monoclonal [EPR10881] to FOXA1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human FOXA1 aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: P55317

  • Positive control
    • WB: SW480 and HepG2 cell lysate. Mouse and rat lung lysates IHC-P: Human breast carcinoma & prostate tissue, mouse liver, Rat pancreas tissue. ICC/IF: PC-3 and HepG2 cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab170933 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 49 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use 1/100 - 1/250

ICC/IF 1/100 - 1/250.
Flow Cyt Use at an assay dependent concentration.

Target

  • Function
    Transcription factor that is involved in embryonic development, establishment of tissue-specific gene expression and regulation of gene expression in differentiated tissues. Is thought to act as a 'pioneer' factor opening the compacted chromatin for other proteins through interactions with nucleosomal core histones and thereby replacing linker histones at target enhancer and/or promoter sites. Binds DNA with the consensus sequence 5'-[AC]A[AT]T[AG]TT[GT][AG][CT]T[CT]-3' (By similarity). Proposed to play a role in translating the epigenetic signatures into cell type-specific enhancer-driven transcriptional programs. Its differential recruitment to chromatin is dependent on distribution of histone H3 methylated at 'Lys-5' (H3K4me2) in estrogen-regulated genes. Involved in the development of multiple endoderm-derived organ systems such as liver, pancreas, lung and prostate; FOXA1 and FOXA2 seem to have at least in part redundant roles (By similarity). Modulates the transcriptional activity of nuclear hormone receptors. Is involved in ESR1-mediated transcription; required for ESR1 binding to the NKX2-1 promoter in breast cancer cells; binds to the RPRM promter and is required for the estrogen-induced repression of RPRM. Involved in regulation of apoptosis by inhibiting the expression of BCL2. Involved in cell cycle regulation by activating expression of CDKN1B, alone or in conjunction with BRCA1. Originally discribed as a transcription activator for a number of liver genes such as AFP, albumin, tyrosine aminotransferase, PEPCK, etc. Interacts with the cis-acting regulatory regions of these genes. Involved in glucose homeostasis.
  • Tissue specificity
    Highly expressed in prostate and ESR1-positive breast tumors. Overexpressed in esophageal and lung adenocarcinomas.
  • Sequence similarities
    Contains 1 fork-head DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • forkhead box A1 antibody
    • Forkhead box protein A1 antibody
    • FOX A1 antibody
    • FOXA1 antibody
    • FOXA1_HUMAN antibody
    • hepatocyte nuclear factor 3 alpha antibody
    • Hepatocyte nuclear factor 3-alpha antibody
    • HNF 3A antibody
    • HNF-3-alpha antibody
    • HNF-3A antibody
    • HNF3A antibody
    • MGC33105 antibody
    • TCF 3A antibody
    • TCF-3A antibody
    • TCF3A antibody
    • Transcription factor 3A antibody
    see all

Images

  • 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.
  • 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 Perform heat mediated antigen retrieval 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
  • 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 Perform heat mediated antigen retrieval 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
  • 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 Perform heat mediated antigen retrieval 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.
  • All lanes : Anti-FOXA1 antibody [EPR10881] (ab170933) at 1.1 µg/ml (purified)

    Lane 1 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Mouse lung lysates
    Lane 3 : Rat lung lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 49 kDa
    Observed band size: 49 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: FOXA1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: MCF7 whole cell lysate (20 µg)
    Lane 4: HepG2 whole cell lysate (20 µg)

    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. 

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

  • Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution.

  • Flow Cytometry analysis of HepG2 (human hepatocellular carcinoma) cells labeling FOXA1 with purified ab170933 at 1/200 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 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.

  • Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling FOXA1 with unpurified ab170933 at 1/100 dilution.

References

This product has been referenced in:
  • Yoshida T  et al. Three-dimensional organoid culture reveals involvement of Wnt/ß-catenin pathway in proliferation of bladder cancer cells. Oncotarget 9:11060-11070 (2018). Read more (PubMed: 29541396) »
  • Ubuka T  et al. Identification of Transmembrane Protease Serine 2 and Forkhead Box A1 As the Potential Bisphenol A Responsive Genes in the Neonatal Male Rat Brain. Front Endocrinol (Lausanne) 9:139 (2018). IHC-P ; Rat . Read more (PubMed: 29643838) »

See all 5 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293T and Prostate cancer cells)
Gel Running Conditions
Reduced Denaturing (4-15% TGX BioRad)
Loading amount
200000 cells
Specification
HEK293T and Prostate cancer cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 50% · Temperature: 25°C
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Submitted Jan 05 2016

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