Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17067] to IRF5 - BSA and Azide free
  • Suitable for: IHC-P, WB, IP, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-IRF5 antibody [EPR17067] - BSA and Azide free
    See all IRF5 primary antibodies

  • Description

    Rabbit monoclonal [EPR17067] to IRF5 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Mouse
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • 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

    Ab231163 is the carrier-free version of ab181553. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab231163 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

  • Western blot - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Western blot - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

  • Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Flow Cytometry - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

  • Immunoprecipitation - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunoprecipitation - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181553).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    This IHC data was generated using the same anti-IRF5 antibody clone, EPR17067, in a different buffer formulation (cat# ab181553).

    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.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Immunocytochemistry/ Immunofluorescence - Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

    This ICC data was generated using the same anti-IRF5 antibody clone, EPR17067, in a different buffer formulation (cat# ab181553).

    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.

     

  • Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)
    Anti-IRF5 antibody [EPR17067] - BSA and Azide free (ab231163)

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