Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IL-18 antibody [EPR19954] (ab207324)

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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
  • Suitable for: Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-IL-18 antibody [EPR19954]
    See all IL-18 primary antibodies

  • Description

    Rabbit monoclonal [EPR19954] to IL-18

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human IL-18 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; HeLa, A431, Jurkat, HEK-293, PC-3 and HaCaT whole cell lysates; Human skin and ovary cancer lysates. Flow Cyt: PC-3 and HaCaT cells.

  • General notes

    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 ab207324 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
Flow Cyt 1/60.
WB 1/200. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).

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] (ab207324)
      Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
      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



      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] (ab207324)
      Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
      All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/200 dilution

      Lane 1 : Wild-type HEK-293 whole cell lysate
      Lane 2 : IL-18 knockout HEK-293 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.

    • Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
      Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
      Anti-IL-18 antibody [EPR19954] (ab207324) at 1/1000 dilution + Human IL-18 active protein at 0.02 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 22 kDa
      Observed band size: 17 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
      Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
      All lanes : Anti-IL-18 antibody [EPR19954] (ab207324) at 1/1000 dilution

      Lane 1 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
      Lane 2 : HaCaT (Human keratinocyte cell line) whole cell lysate
      Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

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


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The WB profiles are consistent with the literature (PMID 12918059; PMID 11470273).

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

      Lane 1 : Human skin lysate
      Lane 2 : Human ovary cancer lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/5000 dilution

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


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Detection in skin and ovary required high concentration of antibody.

    • Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)
      Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)

      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.

    • Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)
      Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)

      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.

    • Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)
      Flow Cytometry - Anti-IL-18 antibody [EPR19954] (ab207324)

      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.

    • Anti-IL-18 antibody [EPR19954] (ab207324)
      Anti-IL-18 antibody [EPR19954] (ab207324)

    References (3)

    Publishing research using ab207324? Please let us know so that we can cite the reference in this datasheet.

    ab207324 has been referenced in 3 publications.

    • Ye Y  et al. Meisoindigo Protects Against Focal Cerebral Ischemia-Reperfusion Injury by Inhibiting NLRP3 Inflammasome Activation and Regulating Microglia/Macrophage Polarization via TLR4/NF-?B Signaling Pathway. Front Cell Neurosci 13:553 (2019). PubMed: 31920554
    • Clement M  et al. MARK4 (Microtubule Affinity-Regulating Kinase 4)-Dependent Inflammasome Activation Promotes Atherosclerosis-Brief Report. Arterioscler Thromb Vasc Biol 39:1645-1651 (2019). PubMed: 31167564
    • Ma ZZ  et al. Expression and clinical significance of the NEK7-NLRP3 inflammasome signaling pathway in patients with systemic lupus erythematosus. J Inflamm (Lond) 15:16 (2018). PubMed: 30202244

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