Human IL-33 Antibody

Catalog #: MAB36254 Datasheet

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

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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

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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

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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.

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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

Onda, H. et al. (1999) J. Cereb. Blood Flow Metab. 19:1279.
Baekkevold, E.S. et al. (2003) Am. J. Pathol. 163:69.
Schmitz, J. et al. (2005) Immunity 23:479.
Black, R.A. et al. (1989) J. Biol. Chem. 264:5323.
Xu, D. et al. (1998) J. Exp. Med. 187:787.
Lohning, M. et al. (1998) Proc. Natl. Acad. Sci. USA 95:6930.
Dinarello, C.A. (2005) Immunity 23:461.
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