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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: WB, ICC/IF, IHC-P
  • Reacts with: Mouse, Rat, Human

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 & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data

  • 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.

  • 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

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 lysate
    Lane 2 : Rat liver lysate
    Lane 3 : Human fetal liver lysate
    Lane 4 : Human stomach 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.

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

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