Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16885] to HNF-4-alpha - ChIP Grade
  • Suitable for: ChIP, IHC-P, WB, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade
    See all HNF-4-alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR16885] to HNF-4-alpha - ChIP Grade

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ChIP
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HepG2 and SW480 whole cell lysates; Human fetal liver, colon and fetal kidney lysates; mouse and rat liver lysates. IHC-P: Human liver, Human colon, mouse liver and rat colon tissues. IP: HepG2 whole cell extract. ChIP: HepG2 whole cell extract.

  • 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

  • ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    ChIP - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

    Chromatin was prepared from HepG2 (Human liver hepatocellular carcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab181604 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

    ABCC6_NC1 is negative control

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/10000 dilution

    Lane 1 : HepG2 whole cell lysate
    Lane 2 : HEK293 whole cell lysate
    Lane 3 : Mouse liver lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution

    Predicted band size: 53 kDa



    This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour before being incubated with ab181604 (rabbit monoclonal [EPR16885] to HNF-4-alpha; dilution 1:10000) and loading control ab18058 (mouse monoclonal [SPM227] to Vinculin; dilution 1:10000) at 4°C overnight. Antibody binding was detected with ab216773 (Goat anti-Rabbit IgG H&L (IRDye® 800CW); green; dilution 1:20000) and ab216776 (Goat anti-Mouse IgG H&L (IRDye® 680RD), red; dilution 1:20000) for 1 hour at room temperature before imaging.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution + Full length mouse HNF-4-gamma recombinant protein at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) 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: 53 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution + SW480 (Human colon adenocarcinoma cell line) 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: 53 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human colon lysate
    Lane 3 : Human fetal kidney 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: 53 kDa


    Exposure time: 1 minute


    This antibody can recognize 6 isoforms in human. The predicted MW are 53KDa, 52 KDa, 47KDa, 56 KDa, 49 KDa and 44 KDa.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Western blot - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    All lanes : Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604) at 1/1000 dilution

    Lane 1 : Mouse liver lysate
    Lane 2 : Rat liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 53 kDa


    Exposure time: 15 seconds


    This antibody can recognize 2 isoforms in mouse and rat. The predicted MW are 53KDa and 52 KDa.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human liver is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of Human colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse liver is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

    Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling HNF-4-alpha with ab181604 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on epithelial cells of rat colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Immunoprecipitation - Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

    HNF-4 alpha was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell extract with ab181604 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab181604 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: HepG2 whole cell extract 10 µg (Input).

    Lane 2: ab181604 IP in HepG2 whole cell extract.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181604 in HepG2 whole cell extract.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)
    Anti-HNF-4-alpha antibody [EPR16885] - ChIP Grade (ab181604)

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