Overview

  • Product name
  • Description
    Goat polyclonal to FOXA2
  • Host species
    Goat
  • Specificity
    This antibody is expected to recognise both reported isoforms.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Xenopus laevis
  • Immunogen

    Synthetic peptide: GVYSRPIMNSS, corresponding to amino acids 447-457 of Human FOXA 2.

  • Positive control
    • Human liver lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab5074 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 4 - 8 µg/ml.
ICC/IF 1/50. PubMed: 19564924
WB Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 48 kDa).Can be blocked with Human FOXA2 peptide (ab23065). A nuclear extraction procedure may be required to concentrate the protein in samples.

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). In embryonic development is required for notochord formation. Involved in the development of multiple endoderm-derived organ systems such as the liver, pancreas and lungs; FOXA1 and FOXA2 seem to have at least in part redundant roles. 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; regulates the expression of genes important for glucose sensing in pancreatic beta-cells and glucose homeostasis. Involved in regulation of fat metabolism. Binds to fibrinogen beta promoter and is involved in IL6-induced fibrinogen beta transcriptional activation.
  • Sequence similarities
    Contains 1 fork-head DNA-binding domain.
  • Post-translational
    modifications
    Phosphorylation on Thr-156 abolishes binding to target promoters and subsequent transcription activation upon insulin stimulation.
  • Cellular localization
    Nucleus. Cytoplasm. Shuttles between the nucleus and cytoplasm in a CRM1-dependent manner and in response to insulin signaling via AKT1 is exported from the nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Forkhead box A2 antibody
    • Forkhead box protein A2 antibody
    • FOX A2 antibody
    • foxa2 antibody
    • FOXA2_HUMAN antibody
    • Hepatic nuclear factor 3 beta antibody
    • Hepatocyte nuclear factor 3 antibody
    • Hepatocyte nuclear factor 3 beta antibody
    • Hepatocyte nuclear factor 3-beta antibody
    • HNF 3B antibody
    • HNF-3-beta antibody
    • HNF-3B antibody
    • HNF3B antibody
    • MGC19807 antibody
    • TCF 3B antibody
    • TCF-3B antibody
    • TCF3B antibody
    • Transcription factor 3B antibody
    see all

Images

  • Anti-FOXA2 antibody (ab5074) at 1 µg/ml + Human Liver lysate at 1 µg/ml

    Predicted band size: 48 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



    Primary incubation was 1 hour. Detected by chemiluminescence.

References

This product has been referenced in:
  • Bhanu NV  et al. Histone modification profiling reveals differential signatures associated with human embryonic stem cell self-renewal and differentiation. Proteomics 16:448-58 (2016). Flow Cyt ; Human . Read more (PubMed: 26631989) »
  • Saitoh I  et al. Choice of Feeders Is Important When First Establishing iPSCs Derived From Primarily Cultured Human Deciduous Tooth Dental Pulp Cells. Cell Med 8:9-23 (2015). Read more (PubMed: 26858904) »
See all 9 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Abreviews
Application
Western blot
Loading amount
60 µg
Gel Running Conditions
Reduced Denaturing (8)
Sample
Xenopus laevis Tissue lysate - whole (whole embryo)
Specification
whole embryo
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C

Abcam user community

Verified customer

Submitted Apr 10 2015

Question
Answer

Vielen Dank für Ihren Anruf.

Wie versprochen habe ich alle unsere FOXA2-Antikörper darauf geprüft, welche Isoformen von FOXA2 sie erkennen. Leider sind alle Immunogene in Regionen lokalisiert, die in beiden Isoformen vorkommen (Details siehe jeweiliges Datenblatt), und daher detektieren die Antikörper immer beide Isoformen. Wir haben also leider keinen Antikörper erhältlich, der zwischen den Isoformen unterscheiden kann.

Es tut mir leid, dass ich keine besseren Nachrichten für Sie habe. Ich hoffe, diese Informationen helfen Ihnen dennoch weiter. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. This antibody is yet to be tested in mouse cells. However, I would predict a favourable result given that the immunogen used to raise this antibody has 100% identity with the mouse homolog. You are largely using an approach that I would recommend. However, we have received a good Abreview from a customer using this antibody; albeit against human cells. The customer applied this antibody with favourable results using a dilution of 1/500. Therefore I would like to recommend that you you try applying this antibody at a reduced dilution. Given that you are preparing a tissue lysate I would also like to recommend that you try probing your blot using an antibody against a housekeeping protein such as GAPDH or beta actin. This will confirm the integrity of this lysate preparation as I would certainly expect detection of this protein in liver cells given the mass of protein that you have been loading. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

I'm sorry to hear you are having a problem with ab5074. This antibody has not been tested in HepG2 cells however it has been tested in human liver lysate and as HepG2 cells are liver derived I would suspect they express good levels of FOxa2. However, they may still express lower levels that liver tissue therefore I would like to recommend to do a nuclear extraction of the protein and to run the positive control of liver lysate along your samples. I would also like to recommend to use 5% BSA in TBST to block the membrane and to try blocking for 1 hour only (and incubating the antibody in TBST only). Finally, can I please confirm that you have checked the transfer of protein is adequate and that the secondary antibody used works with other goat primary antibodies (i.e. that it is not damaged). Please let me know if this helps and do not hesitate to contact me again if you still experience problems with those modifications.

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Answer

Thank you for your enquiry and your interest in our products. Ab5074 recognizes Foxa2 which is a transcription activator for a number of liver genes. As the legend of the Western image on the on-line datasheet shows this antibody has been tested for this application with human liver (whole) tissue lysate. Since the expression levels of Foxa2 in liver cells are very high, normally you do not need to prepare nuclear extract. In case, you would plan to use cell lysate from different tissue or cells than perhaps it would be advisable to prepare nuclear extract. We would strongly suggest running human liver lysate as good positive control along with the samples to see if the detection system and the experimental conditions are fine. We hope this information is useful for you.

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