Recombinant
RabMAb

Recombinant Anti-HNF-4-alpha antibody [EPR3648] - BSA and Azide free (ab227997)

Overview

  • Product name

    Anti-HNF-4-alpha antibody [EPR3648] - BSA and Azide free
    See all HNF-4-alpha primary antibodies
  • Description

    Rabbit monoclonal [EPR3648] to HNF-4-alpha - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
    Unsuitable for: IP
  • 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

    Ab227997 is the carrier-free version of ab92378. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab227997 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Applications

Our Abpromise guarantee covers the use of ab227997 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 Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • 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

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92378).

    • 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).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92378).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92378).

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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92378).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92378).

    • This IHC data was generated using the same anti-HNF4 alpha antibody clone, EPR3648, in a different buffer formulation (cat# ab92378).

      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.

    References

    ab227997 has not yet been referenced specifically in any publications.

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