Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IRF2 antibody [EPR4644(2)] (ab124744)

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Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Immunocytochemistry/ Immunofluorescence - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • OI-RD Scanning - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
  • Anti-IRF2 antibody [EPR4644(2)] (ab124744)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4644(2)] to IRF2
  • Suitable for: WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-IRF2 antibody [EPR4644(2)]
    See all IRF2 primary antibodies

  • Description

    Rabbit monoclonal [EPR4644(2)] to IRF2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human IRF2 aa 300 to the C-terminus.
    Database link: P14316

  • Positive control

  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab124744 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 50 kDa (predicted molecular weight: 39 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
ICC/IF
1/500.
Notes
WB
1/1000. Detects a band of approximately 50 kDa (predicted molecular weight: 39 kDa).
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
ICC/IF
1/500.

Target

  • Function

    Specifically binds to the upstream regulatory region of type I IFN and IFN-inducible MHC class I genes (the interferon consensus sequence (ICS)) and represses those genes. Also acts as an activator for several genes including H4 and IL7. Constitutively binds to the ISRE promoter to activate IL7. Involved in cell cycle regulation through binding the site II (HiNF-M) promoter region of H4 and activating transcription during cell growth. Antagonizes IRF1 transcriptional activation.

  • Tissue specificity

    Expressed throughout the epithelium of the colon. Also expressed in lamina propria.

  • Sequence similarities

    Belongs to the IRF family.
    Contains 1 IRF tryptophan pentad repeat DNA-binding domain.

  • Post-translational
    modifications

    Acetylated by CBP/ p300 during cell-growth. Acetylation on Lys-75 is required for stimulation of H4 promoter activity.
    The major sites of sumoylation are Lys-137 and Lys-293. Sumoylation by SUMO1 increases its transcriptional repressor activity on IRF1 and diminishes its ability to activate ISRE and H4 promoter.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • DKFZp686F0244 antibody
      • Interferon regulatory factor 2 antibody
      • IRF 2 antibody
      • IRF-2 antibody
      • IRF2 antibody
      • IRF2_HUMAN antibody
      see all

    Images

    • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: IRF2 knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: CACO2 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab124744 observed at 48 kDa. Red - loading control, ab18058, observed at 130 kDa.

      ab124744 was shown to recognize IRF2 when IRF2 knockout samples were used, along with additional cross-reactive bands. Wild-type and IRF2 knockout samples were subjected to SDS-PAGE.  Ab124744 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 and 1/10000 dilutions respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      All lanes : Anti-IRF2 antibody [EPR4644(2)] (ab124744) at 1/5000 dilution

      Lane 1 : SW480 (Human colorectal adenocarcinoma cell line)
      Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma)
      Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood)

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 39 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue labeling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution was used as the secondary antibody. Nucleus staining on epithelium of human colon was observed. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

    • Immunocytochemistry/ Immunofluorescence - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Immunocytochemistry/ Immunofluorescence - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling IRF2 with ab124744 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cells.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
      The negative controls are as follows:
      -ve control 1: ab124744 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
      -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

       

    • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      All lanes : Anti-IRF2 antibody [EPR4644(2)] (ab124744) at 1/1000 dilution

      Lane 1 : Caco-2 (Human colorectal adenocarcinoma cells)
      Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus)
      Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells)

      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: 39 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cervical cancer tissue labeling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution was used as the secondary antibody. Nucleus staining on tumor cells of human cervix cancer was observed. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

    • Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Anti-IRF2 antibody [EPR4644(2)] (ab124744) at 1/1000 dilution + Human fetal lung lysate at 10 µg

      Secondary
      Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

      Predicted band size: 39 kDa
      Observed band size: 50 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue labeling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution was used as the secondary antibody. Nucleus staining on epithelial cells of mouse colon was observed. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labelling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

    • OI-RD Scanning - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      OI-RD Scanning - Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-IRF2 antibody [EPR4644(2)] (ab124744)
      Anti-IRF2 antibody [EPR4644(2)] (ab124744)

    References (6)

    Publishing research using ab124744? Please let us know so that we can cite the reference in this datasheet.

    ab124744 has been referenced in 6 publications.

    • Liu T  et al. Global characterization and expression analysis of interferon regulatory factors in response to Aeromonas hydrophila challenge in Chinese soft-shelled turtle (Pelodiscus sinensis). Fish Shellfish Immunol 92:821-832 (2019). PubMed: 31299462
    • Mercado N  et al. IRF2 is a master regulator of human keratinocyte stem cell fate. Nat Commun 10:4676 (2019). PubMed: 31611556
    • Liao W  et al. KRAS-IRF2 Axis Drives Immune Suppression and Immune Therapy Resistance in Colorectal Cancer. Cancer Cell 35:559-572.e7 (2019). PubMed: 30905761
    • Verhoeven RJA  et al. NF-?B Signaling Regulates Epstein-Barr Virus BamHI-Q-Driven EBNA1 Expression. Cancers (Basel) 10:N/A (2018). PubMed: 29659505
    • Cheng L  et al. Repression of let-7a cluster prevents adhesion of colorectal cancer cells by enforcing a mesenchymal phenotype in presence of liver inflammation. Cell Death Dis 9:489 (2018). PubMed: 29695839
    • Camicia R  et al. BAL1/ARTD9 represses the anti-proliferative and pro-apoptotic IFN?-STAT1-IRF1-p53 axis in diffuse large B-cell lymphoma. J Cell Sci 126:1969-80 (2013). PubMed: 23487038

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    ChIP abreview for Anti-IRF2 antibody [EPR4644(2)]

     Good
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (A673/TR/shEF1 Ewing sarcoma cell line)
    Negative control
    IgG isotype antibody in A673/TR/shEF1 Ewing sarcoma cell line
    Specification
    A673/TR/shEF1 Ewing sarcoma cell line
    Detection step
    Other
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: methanol-free formaldehyde
    Positive control
    IRF2 antibody in A673/TR/shEF1 Ewing sarcoma cell line
    Read More

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    Submitted Aug 16 2018

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