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  1. Link

    il-18-antibody-epr19954-ab207324.pdf

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Immunology Innate Immunity Cytokines Interleukins
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Validated using a knockout cell lineRecombinantRabMAb

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

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Western blot - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)
  • Anti-IL-18 antibody [EPR19954] (ab207324)

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: WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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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: WB, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Human
  • Immunogen

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

  • 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 (intra): 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19954
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Innate Immunity
    • Cytokines
    • Interleukins
    • Cancer
    • Invasion/microenvironment
    • Angiogenesis
    • Angiogenic growth factors
    • Cancer
    • Tumor immunology
    • Cytokines
    • Interleukins
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

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

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human IL18 knockout HeLa cell line (ab265274)
  • KO cell lysates

    • Human IL18 knockout HeLa cell lysate (ab256952)
  • KO cell pellets

    • Human IL18 knockout HeLa cell pellet (ab278892)
  • Recombinant Protein

    • Recombinant Human IL-18 protein (ab188059)

Applications

The Abpromise guarantee

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

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.
  • Target information above from: UniProt accession Q14116 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3606 Human
    • Omim: 600953 Human
    • SwissProt: Q14116 Human
    • Unigene: 83077 Human
    • 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 (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)
      Flow Cytometry (Intracellular) - Anti-IL-18 antibody [EPR19954] (ab207324)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling IL-18with 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 Fluorr® 488) at 1/2000 dilution was used as the secondary antibody. 

       

       

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

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HaCaT (Human keratinocyte cell line) cells labeling IL-18with 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 Fluorr® 488) at 1/2000 dilution was used as the secondary antibody. 

       

       

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

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling IL-18with 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 Fluorr® 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)

    Protocols

    • Western blot protocols
    • Flow cytometry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (4)

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

    ab207324 has been referenced in 4 publications.

    • Zhu H  et al. Janus Kinase Inhibition Ameliorates Ischemic Stroke Injury and Neuroinflammation Through Reducing NLRP3 Inflammasome Activation via JAK2/STAT3 Pathway Inhibition. Front Immunol 12:714943 (2021). PubMed: 34367186
    • 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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