Human IL-33 Antibody

(8 citations)
(2 Reviews)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-33 in ELISAs and Western blots. In sandwich ELISAs, less than 0.05% cross-reactivity with recombinant mouse (rm) IL‑33 is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human IL-33
    Ser112-Thr270
    Accession # O95760
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human IL‑33 (Catalog # 3625-IL)
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 µg/mL
    Immersion fixed human peripheral blood mononuclear cells treated with PMA and calcium ionomycin
    • Human IL-33 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    0.2-0.8 µg/mL 
    Human IL‑33 Antibody (Catalog # AF3625)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human IL‑33 Biotinylated Antibody (Catalog # BAF3625)
  • ELISA Standard
     
    Recombinant Human IL-33 Protein (Catalog # 3625-IL)
  • Neutralization
    Measured by its ability to neutralize IL‑33-induced proliferation in the D10.G4.1 mouse helper T cell line. The Neutralization Dose (ND50) is typically 0.75-3.0 µg/mL in the presence of 1 ng/mL Recombinant Human IL‑33 and sub-optimal amounts of Mouse CD3 epsilon Monoclonal Antibody.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Cell Proliferation Induced by IL‑33 and Neutralization by Human IL‑33 Antibody. In the presence of sub-optimal amounts of Hamster Anti‑Mouse CD3 epsilon Monoclonal Antibody (Catalog # MAB484), Recom­binant Human IL‑33 (Catalog # 3625-IL) stimulates proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Under these conditions, proliferation elicited by Recombinant Human IL‑33 (1 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3625). The ND50 is typically 0.75-3.0 µg/mL.
Immunohistochemistry
IL‑33 in Human Tonsil. IL‑33 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3625) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: IL-33

IL-33, also known as NF-HEV and DVS 27, is a 30 kDa proinflammatory protein that may also regulate gene transcription (1‑3). DVS 27 was identifed as a gene that is up‑regulated in vasospastic cerebral arteries (1). NF-HEV was described as a nuclear factor that is preferentially expressed in the endothelial cells of high endothelial venules relative to endothelial cells from other tissues (2). IL-33 was identified based on sequence and structural homology with IL-1 family cytokines (3). DVS 27, NF-HEV, and IL-33 share 100% amino acid sequence identity. IL-33 is constitutively expressed in smooth muscle and airway epithelia. It is up‑regulated in arterial smooth muscle, dermal fibroblasts, and keratinocytes following IL-1 alpha or IL-1 beta stimulation (1, 3). Similar to IL-1, IL-33 can be cleaved in vitro by caspase-1, generating an N-terminal fragment that is slightly shorter than the C-terminal fragment (3, 4). The N-terminal portion of full length IL-33 contains a predicted bipartite nuclear localization sequence and a homeodomain-like helix-turn-helix DNA binding domain. By immunofluorescence, full length IL-33 localizes to the nucleus in HUVECs and transfectants (2). The C-terminal fragment, corresponding to mature IL-33, binds and triggers signaling through mast cell IL-1 R4/ST2L, a longtime orphan receptor involved in the augmentation of Th2 cell responses (3, 5-7). A ternary signaling complex is formed by the subsequent association of IL-33 and ST2L with IL-1R AcP (8). Stimulation of Th2 polarized lymphocytes with mature IL-33 in vitro induces IL-5 and IL-13 secretion (3). In vivo administration of mature IL-33 promotes increased production of IL-5, IL-13, IgE, and IgA, as well as splenomegaly and inflammatory infiltration of mucosal tissues (3). Full length and mature human IL-33 share 52‑58% aa sequence identity with mouse and rat IL-33. Human IL-33 shares less than 20% aa sequence identity with other IL-1 family proteins.

  • References:
    1. Onda, H. et al. (1999) J. Cereb. Blood Flow Metab. 19:1279.
    2. Baekkevold, E.S. et al. (2003) Am. J. Pathol. 163:69.
    3. Schmitz, J. et al. (2005) Immunity 23:479.
    4. Black, R.A. et al. (1989) J. Biol. Chem. 264:5323.
    5. Xu, D. et al. (1998) J. Exp. Med. 187:787.
    6. Lohning, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:6930.
    7. Dinarello, C.A. (2005) Immunity 23:461.
    8. Chackerian, A.A. et al. (2007) J. Immunol. 179:2551.
  • Long Name:
    Interleukin 33
  • Entrez Gene IDs:
    90865 (Human); 77125 (Mouse)
  • Alternate Names:
    C9orf26; C9orf26chromosome 9 open reading frame 26 (NF-HEV); DKFZp586H0523; DVS27; DVS27-related protein; IL1F11; IL-1F11; IL33; IL-33; interleukin 33; Interleukin-1 family member 11; interleukin-33; NFHEV; NF-HEV; NF-HEVNFEHEV; Nuclear factor from high endothelial venules; RP11-575C20.2
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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Applications
Sample Type
  1. TLR2 Activation Limits Rhinovirus-Stimulated CXCL-10 by Attenuating IRAK-1-Dependent IL-33 Receptor Signaling in Human Bronchial Epithelial Cells
    J Immunol, 2016;0(0):.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  2. Interleukin-33 and RANK-L Interplay in the Alveolar Bone Loss Associated to Periodontitis
    PLoS ONE, 2016;11(12):e0168080.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. IL-33 signaling contributes to the pathogenesis of myeloproliferative neoplasms.
    Authors: Mager L, Riether C, Schurch C, Banz Y, Wasmer M, Stuber R, Theocharides A, Li X, Xia Y, Saito H, Nakae S, Baerlocher G, Manz M, McCoy K, Macpherson A, Ochsenbein A, Beutler B, Krebs P
    J Clin Invest, 2015;125(7):2579-91.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  4. The role of IL-33 and its receptor ST2 in human nasal epithelium with allergic rhinitis.
    Authors: Kamekura R, Kojima T, Takano K, Go M, Sawada N, Himi T
    Clin. Exp. Allergy, 2012;42(2):218-28.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  5. The immune-modulating cytokine and endogenous Alarmin interleukin-33 is upregulated in skin exposed to inflammatory UVB radiation.
    Authors: Byrne SN, Beaugie C, O'Sullivan C, Leighton S, Halliday GM
    Am. J. Pathol., 2011;179(1):211-22.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Frozen
  6. Expression of interleukin 1-like cytokine interleukin 33 and its receptor complex (ST2L and IL1RAcP) in human pancreatic myofibroblasts.
    Authors: Nishida A, Andoh A, Imaeda H, Inatomi O, Shiomi H, Fujiyama Y
    Gut, 2010;59(4):531-41.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. Inhibition of interleukin-33 signaling attenuates the severity of experimental arthritis.
    Authors: Palmer G, Talabot-Ayer D, Lamacchia C, Toy D, Seemayer CA, Viatte S, Finckh A, Smith DE, Gabay C
    Arthritis Rheum., 2009;60(3):738-49.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  8. Measurement of interleukin-33 (IL-33) and IL-33 receptors (sST2 and ST2L) in patients with rheumatoid arthritis.
    Authors: Hong YS, Moon S, Moon SJ, Joo YB, Joo Y, Jeon C, Jeon CH, Cho ML, Ju JH, Oh HJ, Heo YJ, Heo Y, Park SH, Kim HY, Min JK
    J Korean Med Sci, 0;26(9):1132-9.
    Species: Human
    Sample Type: Synovial Fluid
    Application: ELISA Development
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Average Rating: 5 (Based on 2 reviews)

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We have 2 reviews tested in 1 species: Human.
We have 2 reviews tested in 2 applications: ELISA, Immunocytochemistry/Immunofluorescence.

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Images Ratings Applications Species Reviewed By Date Details
ELISA Human IL-33 Antibody AF3625
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Excellent
 ELISA Human Anonymous 11/09/2017
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ELISA Human IL-33 Antibody AF3625
ELISA: Human IL-33 Antibody [AF3625]

Summary

ApplicationELISA
Sample TestedSerum and Plasma
SpeciesHuman

Other Experimental Details

Other Experimental DetailsAF3625 was used as both the capture and the detection antibody for the sandwich ELISA for IL-33. The immunoassay standard was 3625-IL. Assay had sensitivity of ~1pg/ml. Many healthy human samples were not detectable.
Immunocytochemistry/Immunofluorescence Human IL-33 Antibody AF3625
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Excellent
 ICC/IF Human Anonymous 10/18/2016
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Immunocytochemistry/Immunofluorescence Human IL-33 Antibody AF3625
Immunocytochemistry/Immunofluorescence: Human IL-33 Antibody [AF3625]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedAdult lung
SpeciesHuman

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