Overview

  • Product name
    Anti-IRF5 antibody [EPR17067]
    See all IRF5 primary antibodies
  • Description
    Rabbit monoclonal [EPR17067] to IRF5
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human IRF5 aa 1-200. The exact sequence is proprietary.
    Database link: Q13568-4

  • Positive control
    • WB: IRF5 transfected HEK293T whole cell lysate; Ramos, THP-1, A20 and RAW 264.7 whole cell lysates; Mouse thymus, rat thymus, mouse spleen and rat spleen lysates. IHC-P: Human spleen, Human breast cancer, Human lung cancer, Human tonsil, mouse liver and rat spleen tissues. ICC/IF: RAW 264.7 and THP-1 cells. Flow Cyt: THP-1 cells. IP: RAW 264.7 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 ab181553 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 55 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/1000.
Flow Cyt 1/50.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/150.

Target

  • Involvement in disease
    Genetic variations in IRF5 are associated with susceptibility to inflammatory bowel disease type 14 (IBD14) [MIM:612245]. IBD14 is a chronic, relapsing inflammation of the gastrointestinal tract with a complex etiology. It is subdivided into Crohn disease and ulcerative colitis phenotypes. Crohn disease may affect any part of the gastrointestinal tract from the mouth to the anus, but most frequently it involves the terminal ileum and colon. Bowel inflammation is transmural and discontinuous; it may contain granulomas or be associated with intestinal or perianal fistulas. In contrast, in ulcerative colitis, the inflammation is continuous and limited to rectal and colonic mucosal layers; fistulas and granulomas are not observed. Both diseases include extraintestinal inflammation of the skin, eyes, or joints.
    Genetic variations in IRF5 are associated with susceptibility to systemic lupus erythematosus type 10 (SLEB10) [MIM:612251]. Systemic lupus erythematosus (SLE) is a chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system.
    Genetic variations in IRF5 are a cause of susceptibility to rheumatoid arthritis (RA) [MIM:180300]. It is a systemic inflammatory disease with autoimmune features and a complex genetic component. It primarily affects the joints and is characterized by inflammatory changes in the synovial membranes and articular structures, widespread fibrinoid degeneration of the collagen fibers in mesenchymal tissues, and by atrophy and rarefaction of bony structures.
  • Sequence similarities
    Belongs to the IRF family.
    Contains 1 IRF tryptophan pentad repeat DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Interferon regulatory factor 5 antibody
    • Interferon regulatory factor 5 bone marrow variant antibody
    • IRF 5 antibody
    • IRF-5 antibody
    • Irf5 antibody
    • IRF5_HUMAN antibody
    • SLEB10 antibody
    see all

Images

  • All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : IRF5 knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 55 kDa



    Lanes 1 - 2: Merged signal (red and green). Green - ab181553 observed at 56 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab181553 was shown to specifically react with IRF5 in wild-type HAP1 cells as signal was lost in IRF5 knockout cells. Wild-type and IRF5 knockout samples were subjected to SDS-PAGE. Ab181553 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Anti-IRF5 antibody [EPR17067] (ab181553)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling IRF5 with ab181553 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
    This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized THP-1 (Human monocytic leukemia cells) cells labeling IRF5 with ab181553 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on THP-1 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: ab181553 at 1/1000 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.

     

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cells labeling IRF5 with ab181553 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining on RAW 264.7 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: ab181553 at 1/1000 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.

     

  • All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/5000 dilution

    Lane 1 : Non-transfected HEK293T (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 2 : IRF5 transfected HEK293T (Human epithelial cells from embryonic kidney) whole cell 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: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 1 minute


    Transfected lysate from abcam (Cat#:ab94265).

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-IRF5 antibody [EPR17067] (ab181553) at 1/2000 dilution + Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 10 µg

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

    Predicted band size: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/50000 dilution

    Lane 1 : THP-1 (Human monocytic leukemia cells) whole cell lysate
    Lane 2 : A20 (Mouse reticulum cell sarcoma cell line) whole cell 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: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/2000 dilution

    Lane 1 : Mouse thymus lysate
    Lane 2 : Rat thymus 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: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-IRF5 antibody [EPR17067] (ab181553) at 1/2000 dilution

    Lane 1 : Mouse spleen lysate
    Lane 2 : Rat spleen 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: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Anti-IRF5 antibody [EPR17067] (ab181553) at 1/5000 dilution + RAW 264.7(Mouse macrophage cells transformed with Abelson murine leukemia virus ) whole cell lysate at 10 µg

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

    Predicted band size: 55 kDa
    Observed band size: 55 kDa


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow cytometric analysis of 2% paraformaldehyde-fixed THP-1 (Human monocytic leukemia cells) cells labeling IRF5 with ab181553 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasmic staining on leukocytes of Human spleen 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.

  • Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasmic staining on leukocyte of Human breast cancer stroma 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.

  • Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on cancer cells of Human lung cancer 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.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on kupffer cells of 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.

  • Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling IRF5 with ab181553 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and cytoplasmic staining on lymphocytes of rat spleen 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.

  • IRF5 was immunoprecipitated from 1mg of RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate with ab181553 at 1/150 dilution. Western blot was performed from the immunoprecipitate using ab181553 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: RAW 264.7 whole cell lysate 10 µg (Input).

    Lane 2: ab181553 IP in RAW 264.7 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab181553 in RAW 264.7 whole cell lysate.

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

References

This product has been referenced in:
  • Jiang Y  et al. TIPE2 governs macrophage polarization via negative regulation of mTORC1. Mol Med Rep 17:952-960 (2018). Read more (PubMed: 29115630) »
  • Bai Q  et al. IRF5 is associated with adverse postoperative prognosis of patients with non-metastatic clear cell renal cell carcinoma. Oncotarget 8:44186-44194 (2017). Read more (PubMed: 28562332) »
See all 3 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Abcam has not validated the combination of species/application used in this Abreview.
Application
ChIP
Sample
Mouse Cell lysate - nuclear (LPS-stimulated bone marrow-derived macrophages)
Negative control
non-binding regions, e.g. Arg1 gene body
Specification
LPS-stimulated bone marrow-derived macrophages
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
Positive control
same sample, using different Ab, selected based on previous ChIP-seq data sets

Abcam user community

Verified customer

Submitted May 04 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up