Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
RecombinantRabMAb

Recombinant Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

Reviews (1) Submit a question References (2)
ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
  • Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19265-130] to HNF-4-alpha - ChIP Grade
  • Suitable for: WB, ICC/IF, ChIP, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seq
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade
    See all HNF-4-alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR19265-130] to HNF-4-alpha - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, ChIP, IP, Flow Cyt (Intra), ChIC/CUT&RUN-seqmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human colon and fetal liver tissue lysates; HepG2, Caco-2 and SW480 whole cell lysates. ICC/IF: HepG2 and SW480 cells. Flow Cyt (intra): HepG2 cells. ChIP: HepG2 cells. IP: HepG2 whole cell lysate. ChIC/CUT&RUN-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 ab200142 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
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
ICC/IF
1/100.
ChIP
Use 5 µg for 25 µg of chromatin.
IP
1/30.
Flow Cyt (Intra)
1/600.
ChIC/CUT&RUN-seq
Use at an assay dependent concentration.

5 µg
Notes
WB
1/1000. Detects a band of approximately 53 kDa (predicted molecular weight: 53 kDa).
ICC/IF
1/100.
ChIP
Use 5 µg for 25 µg of chromatin.
IP
1/30.
Flow Cyt (Intra)
1/600.
ChIC/CUT&RUN-seq
Use at an assay dependent concentration.

5 µg

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

    • ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      ChIC/CUT&RUN sequencing - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

      ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of ab200142 [EPR19265-130]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

      Additional screenshots of mapped reads can be downloaded here.

      The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

    • ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      ChIP - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

      Chromatin was prepared from HepG2 (human hepatocellular carcinoma epithelial cell) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 5 µg of ab200142 (blue), and 20 µl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach).

      ChIP was performed according to the literature (PMID: 18850323).

    • Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      Western blot - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      All lanes : Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142) at 1/2000 dilution

      Lane 1 : Human colon lysate
      Lane 2 : Human fetal liver lysate
      Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 4 : Caco-2 (human colorectal adenocarcinoma cell line) whole cell lysate
      Lane 5 : SW480 (human colorectal adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/2000 dilution

      Developed using the ECL technique.

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



       

      Exposure times: Lanes 1 and 2: 3 minutes; Lanes 3 and 4: 15 seconds; Lane 5: 3 minutes.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cells labeling HNF-4-alpha with ab200142 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining on HepG2 cell line.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      Flow Cytometry (Intracellular) - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

      Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cell line labeling HNF-4-alpha with ab200142 at 1/600 (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

      HNF-4-alpha was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab200142 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab200142 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
      Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) 10 μg (Input).
      Lane 2: ab200142 IP in HepG2 whole cell lysate (+).
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200142 in HepG2 whole cell lysate (-).
      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 minutes.

    • Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      Immunocytochemistry/ Immunofluorescence - Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (human colorectal adenocarcinoma cell line) cells labeling HNF-4-alpha with ab200142 at 1/100 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). Confocal image showing nuclear staining on SW480 cell line.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)
      Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade (ab200142)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (2)

    Publishing research using ab200142? Please let us know so that we can cite the reference in this datasheet.

    ab200142 has been referenced in 2 publications.

    • Koike H  et al. Engineering human hepato-biliary-pancreatic organoids from pluripotent stem cells. Nat Protoc 16:919-936 (2021). PubMed: 33432231
    • Thompson WL & Takebe T Generation of multi-cellular human liver organoids from pluripotent stem cells. Methods Cell Biol 159:47-68 (2020). PubMed: 32586449

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Application

    Filter by Species

    Filter by Ratings

    Western blot abreview for Anti-HNF-4-alpha antibody [EPR19265-130] - ChIP Grade

     Inconclusive
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Mouse intestinal epithelium)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Mouse intestinal epithelium
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 20 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com