货号 | AF1433-SP |
别名 | BNSP; Bone sialoprotein 1; Eta-1; MGC110940; Nephropontin; OPN; osteopontin; secreted phosphoprotein 1bone sialoprotein I, early T-lymphocyteactivation 1); secreted phosphoprotein-1 (osteopontin, bone sialoprotein); Spp1; SPP-1; SPP1/CALPHA1 fusion; Urinary stone protein; uropontin | 全称 | Secreted Phosphoprotein 1 [BNSP] |
反应种属 | Human |
应用 | Western Blot(0.1 µg/mL) Immunohistochemistry(5-15 µg/mL) |
目标/特异性 | Detects human Osteopontin/OPN in direct ELISAs and Western blots. In direct ELISAs, approximately 8% cross-reactivity with recombinant rat Osteopontin and bovine Osteopontin is observed and less than 5% cross-reactivity with recombinant mouse OPN is observed. |
使用方法 | Western Blot: 0.1 µg/mL Immunohistochemistry: 5-15 µg/mL Neutralization: Measured by its ability to neutralize Osteopontin/OPN-mediated adhesion of the HEK293 human embryonic kidney cell line. Hu, D.D. et al. (1995) J. Biol. Chem. 270:26232. The Neutralization Dose (ND50) is typically 2-6 µg/mL in the presence of 1 µg/mL Recombinant Human Osteopontin/OPN. |
来源 | Polyclonal Goat IgG |
产品组分 |
供应商 | R&D Systems |
Entrez Gene IDs | 6696 (Human); 20750 (Mouse); 25353 (Rat); 281499 (Bovine) |
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. Inhibition of Cellular Adhesion by Immunological Targeting of Osteopontin Neoepitopes Generated through Matrix Metalloproteinase and Thrombin Cleavage. | |
纯化方式 | Antigen Affinity-purified |
免疫原 | Human milk-derived Osteopontin/OPN |
内毒素水平 | <0.10 EU per 1 μg of the antibody by the LAL method. |
生物活性 | Human |
标记 | Unconjugated |
溶解方法 | Reconstitute at 0.2 mg/mL in sterile PBS. |
背景 | Osteopontin (OPN, previously also referred to as transformation-associated secreted phosphoprotein, bone sialoprotein I, 2ar, 2B7, early T lymphocyte activation 1 protein, minopotin, calcium oxalate crystal growth inhibitor protein), is a secreted, highly acidic, calcium-binding, RGD-containing, phosphorylated glycoprotein originally isolated from bone matrix (1). Subsequently, OPN has been found in kidney, placenta, blood vessels and various tumor tissues. Many cell types (including macrophages, osteoclasts, activated T cells, fibroblasts, epithelial cells, vascular smooth muscle cells, and natural killer cells) can express OPN in response to activation by cytokines, growth factors or inflammatory mediators. Elevated expression of OPN has also been associated with numerous pathobiological conditions such as atherosclerotic plaques, renal tubulointerstitial fibrosis, granuloma formations in tuberculosis and silicosis, neointimal formation associated with balloon catheterization, metastasizing tumors, and cerebral ischemia. Human OPN cDNA encodes a 314 amino acid (aa) residue precursor protein with a 16 aa residue predicted signal peptide that is cleaved to yield a 298 aa residue mature protein with an integrin binding sequence (RGD), and N- and O-glycosylation sites. By alternative splicing, at least three human OPN isoforms exist. OPN has been shown to bind to different cell types through RGD-mediated interaction with the integrins alpha vbeta1, alpha vbeta3, alpha vbeta5, and non-RGD-mediated interaction with CD44 and the integrins alpha 8beta1 or alpha 9beta1. OPN exists both as a component of extracellular matrix and as a soluble molecule. Functionally, OPN is chemotactic for macrophages, smooth muscle cells, endothelial cells and glial cells. OPN has also been shown to inhibit nitric oxide production and cytotoxicity by activated macrophages. Human, mouse, rat, pig, and bovine OPN share from approximately 40-80% amino acid sequence identity. Osteopontin is a substrate for proteolytic cleavage by thrombin, enterokinase, MMP-3, and MMP-7. The functions of OPN in a variety of cell types were shown to be modified as a result of proteolytic cleavage (2, 3). |
运输条件 | Blue Ice |
存放说明 | 4℃ |
参考文献 |
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Osteopontin/OPN in Human Breast Cancer Tissue. Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using 8 µg/mL Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the surface of epithelial cells in the intralobular duct. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections. | |
Osteopontin/OPN in Human Breast Cancer Tissue. Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) at 15 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). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections. | |
Cell Adhesion Mediated by Osteopontin/OPN and Neutralization by Human Osteopontin/OPN Antibody. Recombinant Human Osteopontin/OPN (Catalog # 1433-OP), immobilized onto a microplate, supports the adhesion of the HEK293 human embryonic kidney cell line in a dose-dependent manner (orange line). Adhesion elicited by Recombinant Human Osteopontin/OPN (1 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433). The ND50 is typically 2‑6 µg/mL. |