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Immunology Innate Immunity Cytokines Interleukins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-IL-18 antibody [EPR19954-188] (ab243091)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)
  • Flow Cytometry - Anti-IL-18 antibody [EPR19954-188] (ab243091)
  • Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091)
  • Anti-IL-18 antibody [EPR19954-188] (ab243091)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19954-188] to IL-18
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-IL-18 antibody [EPR19954-188]
    See all IL-18 primary antibodies
  • Description

    Rabbit monoclonal [EPR19954-188] to IL-18
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human IL-18 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q14116

  • Positive control

    • WB: PC-3, HeLa, A431 and HaCaT whole cell lysates. IHC-P: Human tonsil, liver and kidney tissues. Flow cyt: PC-3 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

  • 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: PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR19954-188
  • 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

Associated products

  • Alternative Versions

    • Anti-IL-18 antibody [EPR19954-188] - BSA and Azide free (ab243295)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • 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)
  • Recombinant Protein

    • Recombinant Human IL-18 protein (ab188059)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

Our Abpromise guarantee covers the use of ab243091 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 1/1000. Detects a band of approximately 22, 14 kDa (predicted molecular weight: 22 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/50.
  • Application notes
    Is unsuitable for ICC/IF.
  • 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 [EPR19954-188] (ab243091)
        Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        All lanes : Anti-IL-18 antibody [EPR19954-188] (ab243091) at 1/1000 dilution

        Lane 1 : Wild-type HeLa cell lysate
        Lane 2 : IL-18 knockout HeLa cell lysate
        Lane 3 : A431 cell lysate
        Lane 4 : Jurkat cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution

        Performed under reducing conditions.

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



        Lanes 1-4: Merged signal (red and green). Green - ab243091 observed at 22 kDa. Red - loading control ab8245 observed at 37 kDa.  

         ab243091 Anti-IL-18 antibody [EPR19954-188] 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. ab243091 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) 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.

         

         

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)

        Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling IL-18 with ab243091 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on distal convoluted tubules of human kidney (PMID: 17687255). Counter stained with hematoxylin.

        Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

        Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)

        Immunohistochemical analysis of paraffin-embedded human liver tissue labeling IL-18 with ab243091 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on Kupffer cells in human liver (PMID: 19084941). Counter stained with hematoxylin.

        Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

        Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-18 antibody [EPR19954-188] (ab243091)

        Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling IL-18 with ab243091 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on subsets of macrophages in tonsil (PMID: 28842466). Counter stained with hematoxylin.

        Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

        Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

      • Flow Cytometry - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        Flow Cytometry - Anti-IL-18 antibody [EPR19954-188] (ab243091)

        Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) (left panel) and PC-3 (human prostate adenocarcinoma cell line) (right panel) cells labeling IL-18 with ab243091 at 1/50 dilution (red) compared with a 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 H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

        Negative control: Jurkat (PMID 15086390).

      • Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        Western blot - Anti-IL-18 antibody [EPR19954-188] (ab243091)
        All lanes : Anti-IL-18 antibody [EPR19954-188] (ab243091) at 1/1000 dilution

        Lane 1 : PC-3 (human prostate adenocarcinoma cell line) whole cell lysate
        Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
        Lane 3 : HaCaT (Human keratinocyte cell line) whole cell lysate

        Lysates/proteins at 20 µg per lane.

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

        Predicted band size: 22 kDa
        Observed band size: 14,22 kDa
        why is the actual band size different from the predicted?



        Exposure time :  Lanes 1 & 2: 3 minutes. Lane 3: 26 seconds.

        Blocking and dilution buffer: 5% NFDM/TBST. 

        The bands observed are consistent with what has been described in the literature. Mature IL-18 protein is detected as a band at 18 kDa, whereas the 14-kDa band represents
        the inactive form (PMID: 15326478).

      • Anti-IL-18 antibody [EPR19954-188] (ab243091)
        Anti-IL-18 antibody [EPR19954-188] (ab243091)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      Datasheets and documents

      • Datasheet
      • SDS

      Certificate of Compliance

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

    • References (0)

      Publishing research using ab243091? Please let us know so that we can cite the reference in this datasheet.

      ab243091 has not yet been referenced specifically in any publications.

      Customer reviews and Q&As

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