货号 | AF3626-SP |
别名 | C9orf26; C9orf26chromosome 9 open reading frame 26 (NF-HEV); DKFZp586H0523; DVS27; DVS27-related protein; IL1F11; IL-1F11; 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 | 全称 | Interleukin 33 |
反应种属 | Mouse |
应用 | Western Blot,Immunohistochemistry,CyTOF-ready,Immunocytochemistry,Intracellular Staining by Flow Cytometry,ELISA Capture (Matched Antibody Pair),ELISA Detection (Matched Antibody Pair),ELISA Standard ,Neutralization |
目标/特异性 | Detects mouse IL-33 in ELISAs and Western blots. In sandwich immunoassays, less than 2% cross-reactivity with recombinant human IL‑33 is observed. |
使用方法 | Western Blot: 0.4 µg/mL Immunohistochemistry: 5-15 µg/mL Immunocytochemistry: 5-15 µg/mL Intracellular Staining by Flow Cytometry: 2.5 µg/106cells ELISA Capture (Matched Antibody Pair): 0.2-0.8 µg/mL ELISA Detection (Matched Antibody Pair): 0.1-0.4 µg/mL ELISA Standard : 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 10-50 ng/mL in the presence of 0.25 ng/mL Recombinant Mouse IL‑33. |
来源 | Polyclonal Goat IgG |
产品组分 |
供应商 | R&D Systems |
Entrez Gene IDs | 90865 (Human); 77125 (Mouse) |
应用文献 | |
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. Aspirin-Exacerbated Respiratory Disease Involves a Cysteinyl Leukotriene-Driven IL-33-Mediated Mast Cell Activation Pathway. | |
纯化方式 | Antigen Affinity-purified |
免疫原 | E. coli-derived recombinant mouse IL-33 Ser109-Ile266 Accession # Q8BVZ5 |
内毒素水平 | <0.10 EU per 1 μg of the antibody by the LAL method. |
生物活性 | Mouse |
标记 | Unconjugated |
溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
背景 | 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 upregulated 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 upregulated 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 mouse IL-33 share approximately 55% and 90% aa sequence identity with human and rat IL-33, respectively. Mouse IL-33 shares less than 25% aa sequence identity with other IL-1 family proteins. |
运输条件 | Blue Ice |
存放说明 | 4℃ |
参考文献 |
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Detection of Mouse IL‑33 by Western Blot. Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with full length mouse IL-33. PVDF membrane was probed with 0.4 µg/mL of Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for IL‑33 at approximately 37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1. | |
IL‑33 in Mouse Spleen. IL‑33 was detected in immersion fixed frozen sections of mouse spleen using Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626) at 5 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 cytoplasm in lymphocytes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections. | |
Detection of IL‑33 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were treated for 5 hours with 50 ng/mL PMA and 1 μg/mL Ca2+ ionomycin then stained with Goat Anti-Mouse IL‑33 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3626, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin. | |
IL‑33 in bEnd.3 Mouse Cell Line. IL‑33 was detected in immersion fixed bEnd.3 mouse endothelioma cell line using Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red, upper panel; Catalog # NL001) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent ICC Staining of Cells on Coverslips. | |
Proliferation Induced by IL‑33 and Neutralization by Human IL‑33 Antibody. Recombinant Mouse IL‑33 (Catalog # 3626-ML) 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). Proliferation elicited by Recombinant Mouse IL‑33 (0.25 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Mouse IL‑33 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3626). The ND50 is typically 10-50 ng/mL. |