Recombinant
RabMAb

Recombinant Anti-IL-33 antibody [EPR17831] (ab187060)

Overview

  • Product name
    Anti-IL-33 antibody [EPR17831]
    See all IL-33 primary antibodies
  • Description
    Rabbit monoclonal [EPR17831] to IL-33
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse IL-33 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: Q8BVZ5

  • Positive control
    • WB: Rat and mouse lung tissue lysate; RAW 264.7 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 ug/ml lipopolysaccharide (LPS) for 24 hours, whole cell lysate. IHC-P: Mouse and rat spleen tissue. ICC/IF: RAW 264.7 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 ug/ml lipopolysaccharide (LPS) for 24 hours cells. FC: RAW 264.7 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 ab187060 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 1/500.
WB 1/1000. Detects a band of approximately 33 kDa (predicted molecular weight: 30 kDa).
IHC-P 1/2000.

Antigen retrieval performed using Universal HIER antigen retrieval reagent (10X) (ab208572).

ICC/IF 1/500.
  • Application notes
    Is unsuitable for IP.
  • 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

    • All lanes : Anti-IL-33 antibody [EPR17831] (ab187060) at 1/1000 dilution

      Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
      Lane 2 : RAW 264.7 treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 ug/ml lipopolysaccharide (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)

      Developed using the ECL technique.

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



      Blocking/Dilution: 5% NFDM/TBST

      Exposure: 3 minutes

      IL-33 expression is induced by LPS treatment of PMA-differentiated RAW 264.7 cells (PMID 19559631; PMID 19933859).

    • All lanes : Anti-IL-33 antibody [EPR17831] (ab187060) at 1/1000 dilution

      Lane 1 : Rat lung tissue lysate at 20 µg
      Lane 2 : Mouse lung tissue lysate at 10 µg

      Secondary
      Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lane 2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Developed using the ECL technique.

      Predicted band size: 30 kDa
      Observed band size: 30 kDa



      Blocking/Dilution: 5% NFDM/TBST

      Exposure: 3 minutes

    • Flow Cytometry analysis of RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 50nM PMA and 5μg/ml LPS for 24h (Red) / Untreated control (Green) labeling IL-33 with ab187060 at 1/500 dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilised with 0.1% Tween-20. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling IL33 with ab187060 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in endothelial cells of mouse spleen is observed (PMID: 12819012). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.

    • Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling IL33 with ab187060 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use. Nuclear staining in endothelial cells of rat spleen is observed (PMID: 12819012). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101), ready to use.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling IL33 with ab187060 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased nuclear staining in RAW 264.7 cells treated with 50 nM Phorbol-12-myristate-13-acetate (PMA) and 5 µg/ml Lipopolysaccharide for 24h.

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    References

    ab187060 has not yet been referenced specifically in any publications.

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