Recombinant
RabMAb

Recombinant Anti-IL-33 antibody [EPR20417] (ab207737)

Overview

  • Product name
    Anti-IL-33 antibody [EPR20417]
    See all IL-33 primary antibodies
  • Description
    Rabbit monoclonal [EPR20417] to IL-33
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Recombinant fragment within Human IL-33 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: O95760

  • Positive control
    • WB: Human IL33 recombinant protein. THP-1 treated with Phorbol-12-myristate-13-acetate and LPS lysate. Human artery, lung, tonsil, stomach and colon lysate. His-tagged rat IL33 recombinant protein. Rat spleen, kidney and lung lysate. IHC-P: Rat lung and spleen tissue. Human tonsil tissue and kidney cancer tissue. ICC/IF: THP-1 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab207737 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. Predicted molecular weight: 30 kDa.
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Under our testing conditions we observed non-specific staining in some human tissues, customers are recommended to optimize IHC testing conditions.

ICC/IF 1/500.

Target

  • Function
    Cytokine that binds to and signals through the IL1RL1/ST2 receptor which in turn activates NF-kappa-B and MAPK signaling pathways in target cells (PubMed:16286016). Involved in the maturation of Th2 cells inducing the secretion of T-helper type 2-associated cytokines. Also involved in activation of mast cells, basophils, eosinophils and natural killer cells. Acts as a chemoattractant for Th2 cells, and may function as an "alarmin", that amplifies immune responses during tissue injury (PubMed:17853410, PubMed:18836528).
    In quiescent endothelia the uncleaved form is constitutively and abundantly expressed, and acts as a chromatin-associated nuclear factor with transcriptional repressor properties, it may sequester nuclear NF-kappaB/RELA, lowering expression of its targets (PubMed:21734074). This form is rapidely lost upon angiogenic or proinflammatory activation (PubMed:18787100).
  • Tissue specificity
    Expressed at high level in high endothelial venules found in tonsils, Peyer patches and mesenteric lymph nodes. Almost undetectable in placenta.
  • Sequence similarities
    Belongs to the IL-1 family. Highly divergent.
  • Domain
    The homeodomain-like HTH domain mediates nuclear localization and heterochromatin association.
  • Post-translational
    modifications
    The full length protein can be released from cells and is able to signal via the IL1RL1/ST2 receptor. However, proteolytic processing by CSTG/cathepsin G and ELANE/neutrophil elastase produces C-terminal peptides that are more active than the unprocessed full length protein. May also be proteolytically processed by calpains (PubMed:19596270). Proteolytic cleavage mediated by apoptotic caspases including CASP3 and CASP7 results in IL33 inactivation (PubMed:19559631). In vitro proteolytic cleavage by CASP1 was reported (PubMed:16286016) but could not be confirmed in vivo (PubMed:19465481) suggesting that IL33 is probably not a direct substrate for that caspase.
  • Cellular localization
    Nucleus. Chromosome. Cytoplasmic vesicle, secretory vesicle. Secreted. Associates with heterochromatin and mitotic chromosomes (PubMed:17185418).
  • Information by UniProt
  • Database links
  • Alternative names
    • C9orf26 antibody
    • CHROMOSOME 9 OPEN READING FRAME 26 antibody
    • DKFZp586H0523 antibody
    • DVS27 antibody
    • DVS27 related protein antibody
    • IL 1F11 antibody
    • IL 33 antibody
    • IL-1F11 antibody
    • IL-33 antibody
    • IL1F11 antibody
    • IL33 antibody
    • IL33_HUMAN antibody
    • Interleukin 1 family member 11 antibody
    • Interleukin 33 antibody
    • INTERLEUKIN 33 NFHEV antibody
    • Interleukin 33 precursor antibody
    • Interleukin-1 family member 11 antibody
    • Interleukin-33 (109-270) antibody
    • Interleukin33 antibody
    • NF HEV antibody
    • NF-HEV antibody
    • NFEHEV antibody
    • NFHEV antibody
    • Nuclear factor for high endothelial venules antibody
    • Nuclear factor from high endothelial venules antibody
    • OTTHUMP00000021041 antibody
    • RP11 575C20.2 antibody
    see all

Images

  • Anti-IL-33 antibody [EPR20417] (ab207737) at 1/1000 dilution + human IL33 recombinant protein (aa112-270), 10 ng

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

    Predicted band size: 30 kDa
    Observed band size: 18 kDa
    why is the actual band size different from the predicted?


    Exposure time: 2 seconds


    Blocking: 5% NFDM/TBST

  • All lanes : Anti-IL-33 antibody [EPR20417] (ab207737) at 1/1000 dilution

    Lane 1 : Untreated THP-1 (human monocytic leukemia monocyte), whole cell lysate
    Lane 2 : THP-1 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA, ab120297) and 5 ug/ml lipopolysaccharides (LPS) for 24 hours, 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: 30 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    The level of IL33 expression can be induced by PMA and LPS treatment (PMID: 19559631).

    Blocking : 5% NFDM/TBST

     

  • All lanes : Anti-IL-33 antibody [EPR20417] (ab207737) at 1/1000 dilution

    Lane 1 : Human artery
    Lane 2 : Human lung
    Lane 3 : Human tonsil
    Lane 4 : Human stomach
    Lane 5 : Human colon

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

    Predicted band size: 30 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?



    Blocking: 5% NFDM/TBST 

    Exposure times:

    Lanes 1-3,5: 3 minutes

    Lanes 4: 30 seconds

  • Anti-IL-33 antibody [EPR20417] (ab207737) at 1/1000 dilution + Recombinant Rat IL-33 protein (ab207126)

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

    Predicted band size: 30 kDa
    Observed band size: 20 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes
  • All lanes : Anti-IL-33 antibody [EPR20417] (ab207737) at 1/1000 dilution

    Lane 1 : Rat spleen
    Lane 2 : Rat kidney
    Lane 3 : Rat lung

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 30 kDa
    Observed band size: 33 kDa why is the actual band size different from the predicted?



    Blocking: 5% NFDM/TBST 

    Exposure times:

    Lane 1: 3 seconds

    Lane 2: 10 seconds

    Lane3: 5 seconds

  • Immunohistochemical analysis of paraffin embedded rat spleen tissue labeling IL33 with ab207737 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on endothelium cells of rat spleen (PMID: 12819012, PMID: 18836528) is observed. 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.

  • Immunohistochemical analysis of paraffin embedded rat lung tissue labeling IL33 with ab207737 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on epithelium and endothelium cells of rat lung (PMID: 18836528) is observed. 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.

  • Immunohistochemical analysis of paraffin embedded human tonsil tissue labeling IL33 with ab207737 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on high endothelial venules of human tonsil (PMID: 12819012) is observed. 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.

  • Immunohistochemical analysis of paraffin embedded human kidney cancer tissue labeling IL33 with ab207737 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on endothelial cells of human kidney cancer (PMID: 12819012) is observed. 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.

  • Immunofluorescent analysis of 4% paraformaldehyde fixed, 0.1% Triton X-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labeling IL33 with ab207737 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (green). Confocal image showing increased nuclear staining on THP-1 cells treated with 50 nM Phorbol-12-myristate-13-acetate (PMA, ab120297) and 5 µg/ml lipopolysaccharides (LPS) for 24 hours. Nuclear counterstain DAPI used at a 1/200 dilution.

    The negative controls are as follows: AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) only on treated and un-treated cells.

References

ab207737 has not yet been referenced specifically in any publications.

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