Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19954] to IL-18 - BSA and Azide free
  • Suitable for: WB, Flow Cyt
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-IL-18 antibody [EPR19954] - BSA and Azide free
    See all IL-18 primary antibodies

  • Description

    Rabbit monoclonal [EPR19954] to IL-18 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment aa 1 to the C-terminus. The exact sequence is proprietary. Corresponds to the mature protein minus the propeptide.
    Database link: Q14116

  • Positive control

    • WB: Human IL-18 active protein; PC-3, Jurkat, A431, HeLa, HEK-293, and HaCaT cell lysates; Human skin and ovary cancer tissue lysates. Flow Cyt: PC-3 and HaCaT cells.

  • General notes

    ab222926 is the carrier-free version of ab207324. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab222926 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    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

Applications

Our Abpromise guarantee covers the use of ab222926 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 Use at an assay dependent concentration. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Augments natural killer cell activity in spleen cells and stimulates interferon gamma production in T-helper type I cells.

  • Sequence similarities

    Belongs to the IL-1 family.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    • Alternative names

      • Iboctadekin antibody
      • IFN gamma inducing factor antibody
      • IFN-gamma-inducing factor antibody
      • IGIF antibody
      • IL 1 gamma antibody
      • IL 18 antibody
      • IL 1g antibody
      • IL-1 gamma antibody
      • IL-18 antibody
      • IL1 gamma antibody
      • IL18 antibody
      • IL18 protein antibody
      • IL18_HUMAN antibody
      • IL1F4 antibody
      • IL1g antibody
      • IL1gamma antibody
      • ILIF4 antibody
      • Interferon gamma inducing factor antibody
      • Interferon gamma-inducing factor antibody
      • Interleukin 1 gamma antibody
      • Interleukin 18 (interferon-gamma-inducing factor) antibody
      • Interleukin 18 antibody
      • Interleukin-1 gamma antibody
      • Interleukin-18 antibody
      • Interleukin18 antibody
      • MGC12320 antibody
      see all

    Images

    • Western blot - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      Western blot - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/200 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : IL18 knockout HeLa cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 22 kDa
      Observed band size: 22 kDa



      This data was developed using the same antibody clone in a different buffer formulation (ab207324).

      Lanes 1-4: Merged signal (red and green). Green - ab207324 observed at 22 kDa. Red - loading control ab8245 observed at 37 kDa.  

       ab207324 Anti-IL-18 antibody [EPR19954] was shown to specifically react with IL-18 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265274 (knockout cell lysate ab256952) was used. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. ab207324 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 200 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.

       

       

    • Western blot - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      Western blot - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/200 dilution

      Lane 1 : Wild-type HEK293 whole cell lysate
      Lane 2 : IL-18 knockout HEK293 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 22 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab207324 observed at 22 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab207324 was shown to recognize IL-18 in wild-type HEK293 cells as signal was lost at the expected MW in IL-18 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and IL-18 knockout samples were subjected to SDS-PAGE. Ab207324 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/200 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 (ab207324).

    • Flow Cytometry - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      Flow Cytometry - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)

      Flow cytometric analysis of 4% paraformaldehyde-fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling IL-18 with ab207324 at 1/60 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 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 (ab207324).

    • Flow Cytometry - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      Flow Cytometry - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)

      Flow cytometric analysis of 4% paraformaldehyde-fixed HaCaT (Human keratinocyte cell line) cells labeling IL-18 with ab207324 at 1/60 dilution (red) compared with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 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 (ab207324).

    • Flow Cytometry - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      Flow Cytometry - Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)

      Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling IL-18 with ab207324 at 1/60 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 (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

      Negative control:

      Jurkat cells serve as a negative cell line as described in PMID 15086390.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab207324).

    • Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)
      Anti-IL-18 antibody [EPR19954] - BSA and Azide free (ab222926)

    References (0)

    Publishing research using ab222926? Please let us know so that we can cite the reference in this datasheet.

    ab222926 has not yet been referenced specifically in any publications.

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