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RecombinantRabMAb

Recombinant Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)

Submit a review Submit a question References (2)
Western blot - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
  • Western blot - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
  • ChIP-sequencing - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
  • Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16786] to HNF-4-alpha - N-terminal
  • Suitable for: ChIP-sequencing, WB, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 555 Alexa Fluor® 594 Carrier Free HRP

Overview

  • Product name

    Anti-HNF-4-alpha antibody [EPR16786] - N-terminal
    See all HNF-4-alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR16786] to HNF-4-alpha - N-terminal

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ChIP-sequencing, WB, ICC/IF, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse and rat liver lysates. Human fetal liver and Human stomach lysates. HepG2 whole cell lysate. IHC-P: Human hepatocellular carcinoma tissue. ICC/IF: HepG2 cells. ChIP-seq: HepG2 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab199431 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
ChIP-sequencing
Use at an assay dependent concentration.
WB
1/10000. Detects a band of approximately 47, 53 kDa (predicted molecular weight: 53 kDa).
ICC/IF
1/250.
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
ChIP-sequencing
Use at an assay dependent concentration.
WB
1/10000. Detects a band of approximately 47, 53 kDa (predicted molecular weight: 53 kDa).
ICC/IF
1/250.
IHC-P
1/400. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

    see all

  • 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

  • Western blot - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    Western blot - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    All lanes : Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431) at 1/10000 dilution

    Lane 1 : Mouse liver tissue lysate
    Lane 2 : Rat liver tissue lysate
    Lane 3 : Human fetal liver tissue lysate
    Lane 4 : Human stomach tissue 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: 47, 53 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    This antibody can recognize 3 isoforms. The predicted MW are 53KDa, 52 KDa and 47KDa, respectively. 

    Blocking/dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    Western blot - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431) 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: 47, 53 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    This antibody can recognize 3 isoforms. The predicted MW are 53KDa, 52 KDa and 47KDa, respectively.

    Blocking/dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)

    Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling HNF-4-alpha with ab199431 at 1/400 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nucleus staining on Human hepatocellular carcinoma tissue is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma) cells labeling HNF-4-alpha with ab199431 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Nuclear staining on HepG2 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab199431 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • ChIP-sequencing - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    ChIP-sequencing - Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)

    Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma cell line) cells. ChIP was performed with 10^7 HepG2 cells and 8 µg of ab199431 [EPR16786]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
    Additional screenshots of mapped reads can be downloaded here.

  • Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)
    Anti-HNF-4-alpha antibody [EPR16786] - N-terminal (ab199431)

References (2)

Publishing research using ab199431? Please let us know so that we can cite the reference in this datasheet.

ab199431 has been referenced in 2 publications.

  • Yu D  et al. High fat diet-induced oxidative stress blocks hepatocyte nuclear factor 4a and leads to hepatic steatosis in mice. J Cell Physiol 233:4770-4782 (2018). PubMed: 29150932
  • Liu L  et al. Hepatic Tmem30a Deficiency Causes Intrahepatic Cholestasis by Impairing Expression and Localization of Bile Salt Transporters. Am J Pathol 187:2775-2787 (2017). PubMed: 28919113

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