| 货号 | AF1507-SP |
| 别名 | APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; tnfa; tnf-a; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha | 全称 | Tumor Necrosis Factor alpha |
| 反应种属 | Canine |
| 应用 | Western Blot(0.1 µg/mL) Immunocytochemistry(5-15 µg/mL) |
| 目标/特异性 | Detects canine TNF-alpha direct ELISAs and Western blots. In direct ELISAs, approximately 70% cross-reactivity with recombinant human TNF‑ alpha, 50% cross-reactivity with recombinant rhesus macaque TNF-alpha, 35% cross-reactivity with recombinant equine TNF‑ alpha, recombinant cotton rat TNF‑ alpha and recombinant rat TNF-alpha, 25% cross-reactivity with recombinant mouse TNF-alpha, and 10% cross-reactivity with recombinant porcine TNF‑ alpha is observed. |
| 使用方法 | Western Blot: 0.1 µg/mL Immunocytochemistry: 5-15 µg/mL Neutralization: Measured by its ability to neutralize TNF‑ alpha -induced cytotoxicity in the L‑929 mouse fibroblast cell line. Matthews, N. and M.L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 221. The Neutralization Dose (ND50) is typically 0.4-1.6 µg/mL in the presence of 100 ng/mL Recombinant Canine TNF‑ alpha and 1 µg/mL actinomycin D. |
| 来源 | Polyclonal Goat IgG |
| 产品组分 |
| 供应商 | R&D Systems |
| Entrez Gene IDs | 7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit) |
| 纯化方式 | Antigen Affinity-purified |
| 免疫原 | E. coli-derived recombinant canine TNF-alpha Val77-Leu233 Accession # P51742 |
| 内毒素水平 | <0.10 EU per 1 μg of the antibody by the LAL method. |
| 生物活性 | Canine |
| 标记 | Unconjugated |
| 溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
| 背景 | Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, apoptosis, and immune system development. TNF-alpha is produced by a wide variety of immune and epithelial cell types (1, 2). Canine TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD) (3). Within the ECD, canine TNF-alpha shares 84-94% aa sequence identity with equine, feline, human, porcine, and rhesus and 69-77% with bovine, cotton rat, mouse, and rat with TNF-alpha. The 26 kDa type 2 transmembrane protein is assembled intracellularly to form a noncovalently linked homotrimer (4). Ligation of this complex induces reverse signaling that promotes lymphocyte co-stimulation but diminishes monocyte responsiveness (5). Cleavage of membrane bound TNF-alpha by TACE/ADAM17 releases a 55 kDa soluble trimeric form of TNF-alpha (6, 7). TNF-alpha trimers bind the ubiquitous TNF RI and the hematopoietic cell-restricted TNF RII, both of which are also expressed as homotrimers (1, 8). TNF-alpha regulates lymphoid tissue development through control of apoptosis (2). It also promotes inflammatory responses by inducing the activation of vascular endothelial cells and macrophages (2). TNF-alpha is a key cytokine in the development of several inflammatory disorders (9). It contributes to the development of type 2 diabetes through its effects on insulin resistance and fatty acid metabolism (10, 11). |
| 运输条件 | Blue Ice |
| 存放说明 | 4℃ |
| 参考文献 |
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Cytotoxicity Induced by TNF‑ alpha and Neutralization by Canine TNF‑ alpha Antibody. Recombinant Canine TNF‑ alpha (Catalog # 1507‑CT) induces cytotoxicity in the the L‑929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Cytotoxicity elicited by Recombinant Canine TNF‑ alpha (100 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Canine TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1507). The ND50 is typically 0.4-1.6 µg/mL in the presence of the metabolic inhibitor actinomycin D (1 µg/mL). | |
TNF‑ alpha in Canine PBMCs. TNF‑ alpha was detected in immersion fixed canine peripheral blood mononuclear cells (PBMCs) stimulated with PMA and calcium ionomycin using Goat Anti-Canine TNF‑ alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1507) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells. |
