Anti-HNF-4-alpha antibody [K9218] - ChIP Grade (ab41898)

Mouse monoclonal HNF-4-alpha antibody [K9218]. Validated in WB, IP, ELISA, IHC, Flow Cyt, ChIP, ICC/IF and tested in Mouse, Rat, Cow, Human. Cited in 40 publication(s).

Overview

  • Product name
    Anti-HNF-4-alpha antibody [K9218] - ChIP Grade
    See all HNF-4-alpha primary antibodies
  • Description
    Mouse monoclonal [K9218] to HNF-4-alpha - ChIP Grade
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, ELISA, IP, IHC-P, ICC/IF, Flow Cyt, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
    Predicted to work with: Pig
  • Immunogen

    Recombinant fragment corresponding to Human HNF-4-alpha aa 3-49.
    Sequence:

    MADYSAALDPAYTTLEFENVQVLTMGNDTSPSEGTNLNAPNSLGVSAL

  • Positive control
    • Human liver hepatocytes and rat intestine epithelial cell.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 3rd April 2019. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab41898 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. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

ChIP Use at an assay dependent concentration. PubMed: 20463007
EMSA Use at an assay dependent concentration.

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 [K9218] - ChIP Grade (ab41898) at 1 µg/ml + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 53 kDa
    Observed band size: 53 kDa
    Additional bands at: 108 kDa, 37 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes


    This image was generated using the ascites version of the product.

  • ICC/IF image of ab41898 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab41898, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) (ab150113) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    This image was generated using the ascites version of the product.

  • Immunohistochemical analysis of obese mouse colon tissue, staining HNF-4-alpha with ab41898 at 1/200 dilution.

    This image was generated using the ascites version of the product.

  • Overlay histogram showing HepG2 cells stained with ab41898 (red line). The cells were fixed with 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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab41898, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HepG2 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.

    This image was generated using the ascites version of the product.

  • ab41898 staining HNF-4-alpha in human liver hepatocytes (10-20 ug/mL) by Immunohistochemistry, formalin-fixed paraffin embedded sections.

    This image was generated using the ascites version of the product.

  • ab41898 staining HNF-4-alpha in Rat Intestine epithelial cell(10-20 ug/mL) by Immunohistochemistry, formalin-fixed paraffin embedded sections.

    This image was generated using the ascites version of the product.

References

This product has been referenced in:
  • Ma Z  et al. Autophagy promotes hepatic differentiation of hepatic progenitor cells by regulating the Wnt/ß-catenin signaling pathway. J Mol Histol 50:75-90 (2019). Read more (PubMed: 30604254) »
  • Rong X  et al. Human bone marrow mesenchymal stem cells-derived exosomes alleviate liver fibrosis through the Wnt/ß-catenin pathway. Stem Cell Res Ther 10:98 (2019). Read more (PubMed: 30885249) »
See all 48 Publications for this product

Customer reviews and Q&As

1-3 of 3 Q&A

Answer

Thank you for contacting us.


There is no BSA in our product antibody.

Our antibody is collected and refined from mouse ascites by precipitation with ammonium sulfate.

And then the precipitate is resolved in physiological saline with 0.1% sodium azide without BSA.

The concentration of the product is adjusted to 1 mg/ml with the same solution.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us.

We are happy to help customers find the most suitable products for their research purposes; however we are not specialized in this particular field and do not want to recommend a product that may be unsuitable. I have done a precursory search and found two products guaranteed in ICC/IF which may suit your needs. These products are:ab41898 Anti-HNF4 antibody [K9218] (https://www.abcam.com/HNF4-antibody-K9218-ChIP-Grade-ab41898.html) and ab59118Anti-TCF2 antibody (https://www.abcam.com/TCF2-antibody-ab59118.html). While these may be good alternatives we encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.

I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

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Answer

We recently carried out Flow cytometry and updated the datasheets in December. Please Aliquot and store at -20°C long term. Avoid repeated freeze / thaw cycles. Storing the antibody at 4deg for extended period (more than two weeks) could affect the activity of the antibody. I have looked through your protocol and would like to ask you some questions in order to understand your procedure better as well as make a few suggestions to improve your results: 1. How much antibody did you use? 2. Have you tried any optimization of the protocol? If you scroll down on the online datasheet you will see the Flow cytometry testing image for HepG2 cells. Conditions used were: Overlay histogram showing HepG2 cells stained with ab41898 (red line). The cells were fixed with 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 followed by the antibody (ab41898, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HepG2 cells fixed with 4%[araformaldehyde/ permeabilized in 0.1% PBS-Tween used under the same conditions. You need to fix your samples either with MeOH or PFA. Otherwise as the target is nuclear the antibody is not able to enter the cell. HNF4 is not on the plasmamembrane/extracellular, thus without sample fixation you will not detect it.

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