Human IL-33 Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB36254-SP
MAB36254-100
Detection of Human IL‑33 by Western Blot.
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Human IL-33 Antibody Summary

Species Reactivity
Human
Specificity
Detects human IL-33 in direct ELISAs.
Source
Recombinant Monoclonal Goat IgG Clone # 40015D
Purification
Protein A or G purified from cell culture supernatant
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 either lyophilized or 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
1 µg/mL
See below
Immunohistochemistry
0.3-25 µg/mL
See below
Neutralization
Measured by its ability to neutralize IL‑33-induced proliferation in the D10.G4.1 mouse helper T cell line. Schmitz, J. et al. (2005) Immunity 23:479. The Neutralization Dose (ND50) is typically 0.75-3 µg/mL in the presence of 12 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.

Scientific Data

Western Blot Detection of Human IL-33 antibody by Western Blot. View Larger

Detection of Human IL‑33 by Western Blot. Western blot shows lysates of HEK293 human embryonic kidney cell line either mock transfected or transfected with human IL-33. PVDF membrane was probed with 1 µg/mL of Recombinant Goat Anti-Human IL-33 Monoclonal Antibody (Catalog # MAB36254) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for IL-33 at approximately 30 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry IL-33 antibody in Human Tonsil by Immunohistochemistry (IHC-P). View Larger

IL‑33 in Human Tonsil. IL-33 was detected in immersion fixed paraffin-embedded sections of human tonsil using Recombinant Goat Anti-Human IL-33 Monoclonal Antibody (Catalog # MAB36254) at 0.3 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.

Neutralization Cell Proliferation Induced by IL-33 and Neutralization by Human IL-33 Antibody. View Larger

Cell Proliferation Induced by IL-33 and Neutralization by Human IL-33 Antibody. In the presence of sub-optimal amounts of Hamster Anti-Mouse CD3e 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), as measured by Resazurin (Catalog # AR002). Under these conditions, proliferation elicited by Recombinant Human IL-33 (12 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL-33 Monoclonal Antibody (Catalog # MAB36254). The ND50 is typically 0.75-3 µg/mL.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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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

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