Overview

  • Product nameAnti-FOXA1 antibody - ChIP Grade
    See all FOXA1 primary antibodies
  • Description
    Goat polyclonal to FOXA1 - ChIP Grade
  • Tested applicationsSuitable for: IHC-P, ChIP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Cow, Pig, Xenopus laevis, Zebrafish
  • Immunogen

    Synthetic peptide:

    C-GVYSRPVLNTS

    , corresponding to amino acids 463-473 of Human FOXA1(Peptide available as ab23074.)

  • Positive control
  • General notes

    Our customers have provided mixed feedback about the Chip application. The data suggests some lots worked perfect but some completely failed. We will be happy to answer any question from potential users. Customers can contact scientific support for latest information about the lot specific data.

Properties

Applications

Our Abpromise guarantee covers the use of ab5089 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
IHC-P Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. PubMed: 20018680
ChIP Use at an assay dependent concentration.
WB Use a concentration of 0.3 - 1 µg/ml. Detects a band of approximately 50 kDa (predicted molecular weight: 49.1 kDa).Can be blocked with Human FOXA1 peptide (ab23074).

An additional band of unknown identity was also consistently observed at 37kDa. This band was successfully blocked by incubation with the immunizing peptide.

Target

  • FunctionTranscription 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 specificityHighly expressed in prostate and ESR1-positive breast tumors. Overexpressed in esophageal and lung adenocarcinomas.
  • Sequence similaritiesContains 1 fork-head DNA-binding domain.
  • Cellular localizationNucleus.
  • 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

Anti-FOXA1 antibody - ChIP Grade images

  • Goat polyclonal to FOXA1 antibody (ab5089) was used in a ChIP assay to detect in vivo binding of FOXA1/HNF3a to the promoter of the TFF1 gene in MCF-7 cells. The cells were fixed in 1% formaldehyde for 15 minutes at room temperature and ChIP was performed using 6µg/ml ab5089. Real-time PCR was used to quantify the enrichment of the TFF1 promoter in the FOXA1 ChIP'd DNA, using the input DNA as the reference. The results indicate that there is an 18-fold enrichment of the TFF1 promoter in the DNA ChIp'd with ab5089.  

    This image was submitted as part of a review.

    See Abreview

  • Chromatin was prepared from nuclear lysate of the human MCF7 breast epithelial adenocarcinoma cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in formaldehyde. The primary antibody was diluted to 0.2 μg/μg chromatin and incubated with the sample for 16 hours at 4°C in a dilution buffer that contains SDS, DOC, Triton X-100, EDTA, HEPES, NaCl. The immunoprecipitated DNA was quantified by real time PCR. Ct values were converted to DNA copy numbers using a standard curve in the Q-PCR step. The number of binding events detected for each test reaction was then calculated by taking into account the DNA copy number, cell equivalents of chromatin used in the ChIP and PCR, and primer pair amplification efficiency.

    See Abreview

  • ab5089 at 5 µg/ml staining FOXA1 in Human prostate tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Steamed antigen retrieval with citrate buffer pH 6, AP-staining.
  • Anti-FOXA1 antibody - ChIP Grade (ab5089) at 0.3 µg/ml + HepG2 at 35 µg

    Predicted band size : 49.1 kDa
    Observed band size : 50 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 37 kDa. We are unsure as to the identity of these extra bands.

    An additional band of unknown identity was also consistently observed at 37kDa.

References for Anti-FOXA1 antibody - ChIP Grade (ab5089)

This product has been referenced in:
  • Griffon A  et al. Integrative analysis of public ChIP-seq experiments reveals a complex multi-cell regulatory landscape. Nucleic Acids Res 43:e27 (2015). Read more (PubMed: 25477382) »
  • Sasahara M  et al. Overexpression of androgen receptor and forkhead-box A1 protein in apocrine breast carcinoma. Anticancer Res 34:1261-7 (2014). Human . Read more (PubMed: 24596370) »

See all 25 Publications for this product

Product Wall

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The following lot numbers are currently in stock, GR115830-1 and GR115838-1.

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Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products.
To our knowledge, ab5089 has not been tested in immunohistochemistry on frozen tissue sections (IHC-Fr). ...

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Nous sommes désolés d'apprendre que les deux derniers tube de cet anticorps que vous avez reçu ne fonctionnent pas comme attendus. Comme discuté au téléphone, co...

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We have ab5089 and ab115279 antibodies that are predicted to react based on sequence homology between human and Porcine. These are the best suitable antibodies we can offer and are eligible for 100% Abreview disc...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (MCF7 breast epithelial adenocarcinoma)
Specification MCF7 breast epithelial adenocarcinoma
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step Real-time PCR
Positive control Region 3' of the CCND1 gene
Region upstream of the transcription start site of the TFF1 gene
Negative control Region on chromosome 4 (Untr4) that is far from any known gene annotation and not expected to be bound by FOXA1
Region on chromosome 20 (Untr20) that is far from any known gene annotation and not expected to be bound by FOXA1
Username

Mr. Genpathway Inc

Verified customer

Submitted Jul 03 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - other (HepG2)
Loading amount 1e+006 cells
Specification HepG2
Gel Running Conditions Reduced Denaturing (4-12% NuPAGE)
Blocking step Blocking agent - RPN2125V (GE) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Mr. Mehdi Motallebipour

Verified customer

Submitted May 16 2008

Thank you for your enquiry regarding the specificity of our antibody raised against FOXA1 (ab5089). There is 72% sequence homology between the FOXA1 synthetic peptide immunogen used to raise this antibody and FOXA2. This means there is a small chance ...

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Thank you for your enquiry and patience. I enquired with the originator of this antibody and we cannot find references to FOXA1 isoforms in Entrez Gene or SwissProt. The sequence of the immunogen used to generate this antibody is as follows: GVYSRPVL...

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