Overview

  • Product name
    Anti-FOXA1 antibody - ChIP Grade
    See all FOXA1 primary antibodies
  • Description
    Goat polyclonal to FOXA1 - ChIP Grade
  • Host species
    Goat
  • Tested applications
    Suitable 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 corresponding to Human FOXA1 aa 462-472 (C terminal).
    Sequence:

    C-GVYSRPVLNTS


    Database link: P55317
    (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

  • 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

  • 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

This product has been referenced in:
  • Liu Y  et al. Neuronal IFN-beta-induced PI3K/Akt-FoxA1 signalling is essential for generation of FoxA1(+)Treg cells. Nat Commun 8:14709 (2017). IP ; Mouse . Read more (PubMed: 28436428) »
  • Wong ES  et al. Interplay of cis and trans mechanisms driving transcription factor binding and gene expression evolution. Nat Commun 8:1092 (2017). Read more (PubMed: 29061983) »
See all 52 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Answer

Thank you for contacting us.

The lot numbers you have kindly asked are sold out.

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

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us.
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). However by participating in our AbTrial program you can now use our products in an untested application or species without financial risk.
Simply follow these easy steps below to apply for our AbTrial Program:
1. Reply to this email, letting us know you are interested in testing this product.
2. Our scientists will email you an inactive personal discount code for the value of the product.
3. Purchase and test the product at the regular price.
4. Submit your results, including your discount code in the additional notes section of your Abreview.
5. Once the Abreview is submitted, the discount code will become active.
6. Apply your discount code on your next order to receive that value off.
Please let me know if you have any questions about this offer and I would be happy to help you further.
The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

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Answer

I am sorry this product did not perform as stated on the data sheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab23738 - Anti-FOXA1 antibody.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Answer

Merci de votre émail.

Je suis heureuse d'apprendre que le nouveau lot fonctionne de nouveau comme attendu. Veuillez encore une fois accepter toutes nos excuses pour les problèmes que vous aviez eu avec les deux tubes précédents. Je me permets de vous transférer le numéro de commande du deuxième tube deremplacement gratuit de ce produit:xx. Il devrait arriver demain chez vous. Vous recevrez aussi prochainement un mail de confirmation comprenant les détails d'expédition.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

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Answer

Merci de nous avoir contactés.

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, comme nous avons que le même lot en stock, je vous envoie une unité gratuite de remplacement. Si cette unité vous donne des bons résultats, nous allons vous envoyer un deuxième tube gratuitement. Voicile numéro de commande de remplacement gratuit de ce produit : xx. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

J'espère que le nouveau tube va donner les résultats attendus. Veuillez ne pas hésiter à nous recontacter aussi si vous avec des questions. Je me réjouis d'avoir de vous nouvelles.

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Answer

Thank you for contacting us.

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 discount programme.

Clickthe linkfor more info:

https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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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

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

Mr. Mehdi Motallebipour

Verified customer

Submitted May 16 2008

Answer

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 that it will also detect FOXA2, perhaps giving some weak staining from cross-reactivity. I am sorry that to my knowledge there is no experimental data available on this. I hope this helps. Please get back to me if you have any more questions and I will be happy to help.

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Answer

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: GVYSRPVLNTS, corresponding to amino acids 463-473 of Human FOX A1. So if you have the sequences of the isoforms, you can compare the immunogen sequence with them. Please contact us again if you have any additional questions.

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1-10 of 11 Abreviews or Q&A

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