Recombinant
RabMAb

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

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, Flow Cyt, ChIP, IPmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human HNF-4-alpha aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: P41235

  • 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: HepG2 cells. ChIP: HepG2 cells. IP: HepG2 whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.
Flow Cyt 1/600.
ChIP Use 5 µg for 25 µg of chromatin.
IP 1/30.

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

  • 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).

  • 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.

  • 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 cytometric analysis of 4% 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.

  • 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.

  • 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.

References

ab200142 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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

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Verified customer

Submitted Aug 20 2018

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