|Detection of Bovine IL‑4 by Western Blot. Western blot shows Recombinant Bovine IL‑4 (Catalog # 2469-BL) (10 ng/lane). PVDF membrane was probed with 2 µg/mL of Mouse Anti-Bovine IL‑4 Monoclonal Antibody (Catalog # MAB2469) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for IL‑4 at approximately 12 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|IL‑4 in Bovine PBMCs. IL‑4 was detected in immersion fixed bovine peripheral blood mononuclear cells (PBMCs) stimulated with concanavalin A using Mouse Anti-Bovine IL‑4 Monoclonal Antibody (Catalog # MAB2469) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.|
Interleukin-4 (IL-4), also known as B cell-stimulatory factor-1, is a monomeric, approximately 13 kDa‑18 kDa Th2 cytokine that shows pleiotropic effects during immune responses (1‑3). It is a glycosylated polypeptide that contains three intrachain disulfide bridges and adopts a bundled four alpha -helix structure (4). Bovine IL-4 is synthesized with a 24 amino acid (aa) signal sequence. Alternate splicing generates two additional isoforms with internal deletions (5). Mature bovine IL-4 shares 60%, 91%, 93%, 78%, 55%, 39%, and 41% aa sequence identity with equine, goat, ovine, porcine, human, mouse, and rat IL-4, respectively. IL-4 exerts its effects through two receptor complexes (6, 7). The type I receptor, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R alpha and the common gamma chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and -21). The type II receptor on nonhematopoietic cells consists of IL-4 R alpha and IL-13 R alpha 1. The type II receptor also transduces IL-13 mediated signals. IL-4 is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils (1, 2). It promotes cell proliferation, survival, and immunoglobulin class switch to IgE in B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells (8‑11). IL-4 plays a dominant role in the development of allergic inflammation and asthma (10, 12).
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