|Detection of Human ST2/IL-33 R by Western Blot. Western blot shows lysates of human kidney tissue and human lung tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-HumanST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for ST2/IL-33 R at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Detection of ST2/IL-33 R in KG‑1a Human Cell Line by Flow Cytometry.|
KG‑1a human acute myelogenous leukemia cell line was stained with Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108).
|Detection of Human ST2/IL-33 R by Simple WesternTM. Simple Western lane view shows lysates of human kidney tissue and human lung tissue, loaded at 0.2 mg/mL. A specific band was detected for ST2/IL-33 R at approximately 60 kDa (as indicated) using 2 µg/mL of Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.|
IFN-gamma Secretion Induced by|
IL‑33 and Neutralization by Human ST2/IL-33 R Antibody. Recombinant Human IL‑33 induces IFN-gamma secretion in Human Peripheral Blood Mononuclear cells (PBMC) in the presence of 0.25 ng/mL Recombinant Human IL-12 (Catalog # 219-IL) in a dose-dependent manner (orange line), as measured by the Human IFN-gamma Q-kit (Catalog # DIF50). Under these conditions, IFN-gamma secretion elicited by IL‑33 is neutralized (green line) by increasing concentrations of Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523). The ND50 is typically 0.1-0.6 μg/mL.
ST2, also known as IL-1 R4 and T1, is an Interleukin-1 receptor family glycoprotein that contributes to Th2 immune responses (1, 2). Human ST2 consists of a 310 amino acid (aa) extracellular domain (ECD) with three Ig-like domains, a 21 aa transmembrane segment, and a 207 aa cytoplasmic domain with an intracellular TIR domain (3, 4). Alternate splicing of the 120 kDa human ST2 generates a soluble 60 kDa isoform that lacks the transmembrane and cytoplasmic regions as well as an isoform that additionally lacks the third Ig‑like domain (4). Within the ECD, human ST2 shares 68% and 64% aa sequence identity with mouse and rat ST2, respectively. ST2 is expressed on the surface of mast cells, activated Th2 cells, macrophages, and cardiac myocytes (5‑8). It binds IL-33, a cytokine that is upregulated by inflammation or mechanical strain in smooth muscle cells, airway epithelia, keratinocytes, and cardiac fibroblasts (5, 9). IL-33 binding induces the association of ST2 with IL-1R AcP, a shared signaling subunit that also associates with IL-1 RI and IL-1 R rp2 (1, 10, 11). In macrophages, ST2 interferes with signaling from IL-1 RI and TLR4 by sequestering the adaptor proteins MyD88 and Mal (7). In addition to its role in promoting mast cell and Th2 dependent inflammation, ST2 activation enhances antigen induced hypernociception and protects from atherosclerosis and cardiac hypertrophy (5, 12‑14). The soluble ST2 isoform is released by activated Th2 cells and strained cardiac myocytes and is elevated in the serum in allergic asthma (6, 8, 15). Soluble ST2 functions as a decoy receptor that blocks IL‑33’s ability to signal through transmembrane ST2 (10, 13‑15).
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ELISA: Human ST2/IL-33 R Antibody [AF523]
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