RabMAb

Anti-HNF-4-alpha antibody [EPR3648] (ab92378)

Overview

  • Product nameAnti-HNF-4-alpha antibody [EPR3648]
    See all HNF-4-alpha primary antibodies
  • Description
    Rabbit monoclonal [EPR3648] to HNF-4-alpha
  • Tested applicationsWB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human HNF-4-alpha aa 1-100.

  • Positive control
    • WB: HepG2, A549 and SW480 cell lysates. IHC-P: Human colon and kidney tissues. ICC/IF: HepG2 cells. Flow Cyt: HepG2 cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

Our Abpromise guarantee covers the use of ab92378 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 1/2000. Predicted molecular weight: 53 kDa.

For unpurified use at 1/1000 - 1/10000.

IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/100 - 1/250.

Flow Cyt 1/70.

For unpurified use at 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/100 - 1/250.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionTranscriptionally 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 diseaseDefects 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 similaritiesBelongs 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 localizationNucleus.
    • 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

    Anti-HNF-4-alpha antibody [EPR3648] images

    • Anti-HNF-4-alpha antibody [EPR3648] (ab92378) at 1/2000 dilution (purified) + SW480 cell lysate at 20 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

      Blocking and dilution buffer: 5% NFDM/TBST.

    • Anti-HNF-4-alpha antibody [EPR3648] (ab92378) at 1/5000 dilution (purified) + HepG2 cell lysate at 10 µg

      Secondary
      Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-HNF-4-alpha antibody [EPR3648] (ab92378) at 1/1000 dilution (unpurified)

      Lane 1 : HepG2 cell lysate
      Lane 2 : A549 cell lysate
      Lane 3 : SW480 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size : 53 kDa
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling HNF-4-alpha with purified ab92378 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue (A) and human kidney tissue (B) labelling HNF-4-aplha with unpurified ab92378 at a 1/100 dilution. Detection: DAB staining.

    • Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling HNF-4 with purified ab92378 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    • Immunocytochemistry/Immunfluorescence analysis of HepG2 cells labelling HNF-4-alpha with unpurified ab92378 at a 1/100 dilution.

    • Flow Cytometry analysis of HepG2 cells labelling HNF-4 with purified ab92378 at 1/70 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • Overlay histogram showing HepG2 cells stained with unpurified ab92378 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (unpurified ab92378, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    References for Anti-HNF-4-alpha antibody [EPR3648] (ab92378)

    ab92378 has not yet been referenced specifically in any publications.

    Product Wall

    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (iCell® Cardiomyocytes)
    Permeabilization No
    Specification iCell® Cardiomyocytes
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative Acetone
    Username

    Abcam user community

    Verified customer

    Submitted Aug 11 2015

    Application Flow Cytometry
    Fixation Acetone
    Permeabilization No
    Sample Human Cell (iCell­ Cardiomyocytes)
    Specification iCell­ Cardiomyocytes
    Gating Strategy negative control cells
    Preparation Cell harvesting/tissue preparation method: 1 mM EDTA/1 mM EGTA in PBS (calcium- and magnesium-free)
    Sample buffer: DPBS
    Username

    Abcam user community

    Verified customer

    Submitted Feb 16 2015

    Our antigen retrieval method used for ab92378 and ab129189 is described below.

    Antigen Retrieval

    This is recommended Heat Induced Epitope Retrieval (HIER) using Decloaking Chamber/Pressure Cooker. Hot water bath or Microwave with te...

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    Thank you fo ryour message.

    To clarify regarding ab135618 Anti-PAX8 antibody, the datasheet states we sell this at 100 ug, at a concentration of 0.25 mg/ml. Therefore, you can calculate that at this concentrations, 100 ug will be in 250 ul o...

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    Thank you for your message which has been forwarded to the scientific support team.

    I am sorry it is difficult to let you know the amount of antibody required for one analysis. This will be very individual and will depend on:

    1. T...

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    Thank you for contacting us. Antibodies are viable for 1 year at 4C and 10 years at -20C. Please check the datasheet of individual antibody for storage instructions. I hope this information is helpful to you. Please do not hesita...

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    A BLAST search shows that ab92378 can detect isoforms 1 and 7. This antibody can also detect isoforms 2, 4, and 8. Sequence alignment shows that its immunogen shares 100% identity with isoforms 3 and 9. Overall, it is predicted to react with all isofor...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"