Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2419Y] to IRF3 - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-IRF3 antibody [EP2419Y] - BSA and Azide free
    See all IRF3 primary antibodies

  • Description

    Rabbit monoclonal [EP2419Y] to IRF3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cytmore details
    Unsuitable for: ICC

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • WB: A549, MCF7, HeLa, HAP1, U-937, THP-1,Daudi and Jurkat cell lysates. IP: HeLa whole cell lysate. IHC: Human bladder carcinoma tissue. Flow cyt: HeLa cells.

  • General notes

    ab201810 is the carrier-free version of ab76409 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

    Ab201810 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse, 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    All lanes : Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    Lane 3 : U-937 (Human histiocytic lymphoma monocyte) whole cell lysate
    Lane 4 : THP-1 (Human monocytic leukemia monocyte) whole cell lysate
    Lane 5 : Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 47 kDa



    We are unsure how to define the extra bands.

  • Flow Cytometry - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Flow Cytometry - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    This data was developed using 201810, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling IRF3 with Purified 201810 at 1:20 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    This data was developed using 201810, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue sections labeling IRF3 with Purified 201810 at 1:100 dilution (1.55 µg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) . Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    All lanes : Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : Wild-type HeLa cell lysate
    Lane 4 : IRF3 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab76409).

    Lanes 1 - 4: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control, ab8245 observed at 37 kDa.  

     ab76409 was shown to react with IRF3 in wild-type HeLa. Loss of signal was observed when knockout cell line ab255345 (knockout cell lysate ab263784) was used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab76409 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunoprecipitation - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Immunoprecipitation - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)

    This data was developed using ab76409, the same antibody clone in a different buffer formulation.
    Purified ab76409 at 1/20 dilution (0.8µg) immunoprecipitating IRF3 in HeLa whole cell lysate.
    Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab76409 + HeLa whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76409 in HeLa whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 47 kDa
    Fresh lysate need to be used to avoid protein degradation.

  • Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    All lanes : Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : IRF3 knockout A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 50 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab76409).

    Lanes 1 - 2: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab76409 was shown to react with IRF3 in wild-type A549 cells in western blot with loss of signal observed in IRF3 knockout cell line ab267098 (IRF3 knockout cell lysate ab256954). Wild-type and IRF3 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab76409 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    Western blot - Anti-IRF3 antibody [EP2419Y] - BSA and Azide free (ab201810)
    All lanes : Anti-IRF3 antibody [EP2419Y] (ab76409) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : IRF3 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 47 kDa



    This WB data was generated using the same anti-IRF3 antibody clone, EP2419Y, in a different buffer formulation (cat# ab76409).

    Lanes 1 - 4: Merged signal (red and green). Green - ab76409 observed at 50 kDa. Red - loading control, ab8245, observed at 37kDa.
    ab76409 was shown to react with IRF3 in wild-type HAP1 cells alond with additional cross-reactive bands. No band was observed when IRF3 knockout samples were used. Wild-type and IRF3 knockout samples were subjected to SDS-PAGE. ab76409 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

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