Recombinant
RabMAb

Recombinant Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460)

Overview

  • Product name
    Anti-HNF-4-alpha antibody [EPR16885-99]
    See all HNF-4-alpha primary antibodies
  • Description
    Rabbit monoclonal [EPR16885-99] to HNF-4-alpha
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Mouse HNF-4-alpha aa 1-200. The exact sequence is proprietary.
    Database link: P49698

  • Positive control
    • WB: HepG2 and Caco-2 whole cell lysates; Human colon and fetal kidney lysates. IHC-P: Human colon, Human liver, mouse liver and rat colon tissues. ICC/IF: HepG2 and HT-29 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab201460 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
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

WB 1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/2000.

Target

  • Function
    Transcriptionally controlled transcription factor. Binds to DNA sites required for the transcription of alpha 1-antitrypsin, apolipoprotein CIII, transthyretin genes and HNF1-alpha. May be essential for development of the liver, kidney and intestine.
  • Involvement in disease
    Defects in HNF4A are the cause of maturity-onset diabetes of the young type 1 (MODY1) [MIM:125850]; also symbolized MODY-1. MODY is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
  • Sequence similarities
    Belongs to the nuclear hormone receptor family. NR2 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Post-translational
    modifications
    Phosphorylated on tyrosine residue(s); phosphorylation is important for its DNA-binding activity. Phosphorylation may directly or indirectly play a regulatory role in the subnuclear distribution.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ39654 antibody
    • FRTS4 antibody
    • Hepatic nuclear factor 4 alpha antibody
    • Hepatocyte nuclear factor 4 alpha antibody
    • Hepatocyte nuclear factor 4 antibody
    • Hepatocyte nuclear factor 4-alpha antibody
    • HNF 4 alpha antibody
    • HNF 4 antibody
    • HNF-4-alpha antibody
    • HNF4 antibody
    • HNF4A antibody
    • HNF4A_HUMAN antibody
    • HNF4a7 antibody
    • HNF4a8 antibody
    • HNF4a9 antibody
    • Hnf4alpha antibody
    • HNF4alpha10/11/12 antibody
    • MODY 1 antibody
    • MODY antibody
    • MODY1 antibody
    • NR2A1 antibody
    • NR2A21 antibody
    • Nuclear receptor subfamily 2 group A member 1 antibody
    • OTTHUMP00000031060 antibody
    • OTTHUMP00000031062 antibody
    • TCF 14 antibody
    • TCF antibody
    • TCF-14 antibody
    • TCF14 antibody
    • Tcf4 antibody
    • Transcription factor 14, hepatic nuclear factor antibody
    • Transcription factor 14 antibody
    • Transcription factor HNF 4 antibody
    • Transcription factor HNF-4 antibody
    • Transcription factor HNF4 antibody
    see all

Images

  • Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution + Full length mouse HNF-4-gamma recombinant protein at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Based on sequence analysis of the immunogen, ab201460 shares 72% homology with HNF4 gamma. The levels of XR were tested in the accompanying recombinant protein, WB experiment.

  • Flow Cytometry analysis of HepG2(human hepatocellular carcinoma) labelling CDKN2A/p16INK4a with purified ab201460 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Alexa Fluor® 488 goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HT-29 (Human colorectal adenocarcinoma cells) cells labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution (green).

    Confocal image showing nuclear staining on HT-29 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is stained with ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
    -ve control 1: ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
    -ve control 2: ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton-X100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution (green).

    Confocal image showing nuclear staining on HT-29 cell line.

    The nuclear counter stain is DAPI (blue). Tubulin is stained with ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution (red).
    -ve control 1: ab201460 at 1/2000 dilution followed by AlexaFluor®594 Goat anti-Mouse secondary antibody (ab150120) at 1/500 dilution.
    -ve control 2: ab7291 anti-Tubulin (mouse mAb) at 1/1000 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/500 dilution.

     

  • Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/10000 dilution + HepG2 (Human liver hepatocellular carcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution + Caco-2 (Human colorectal adenocarcinoma cells) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-HNF-4-alpha antibody [EPR16885-99] (ab201460) at 1/1000 dilution

    Lane 1 : Human colon lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size: 53 kDa
    Observed band size: 53 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on Human colon tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on Human liver tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on mouse liver tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab201460 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

    Nuclear staining on rat colon tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

References

This product has been referenced in:
  • Baciu C  et al. Systematic integrative analysis of gene expression identifies HNF4A as the central gene in pathogenesis of non-alcoholic steatohepatitis. PLoS One 12:e0189223 (2017). Read more (PubMed: 29216278) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Liver)
Permeabilization
Yes - 0.3%TritonX100
Specification
Liver
Blocking step
normal goat serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

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Verified customer

Submitted Oct 09 2018

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