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; PC-3 and HaCaT whole cell lysates; Human skin and ovary cancer lysates. Flow Cyt: PC-3 and HaCaT cells.
  • 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 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

  • 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.

  • 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.

  • 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).

  • 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 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 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 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.

References

This product has been referenced in:

  • 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). Read more (PubMed: 30202244) »
See 1 Publication for this product

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