Human Osteopontin/OPN Antibody Summary
Ile17-Asn300
Accession # NP_000573.1
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Scientific Data
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Detection of Human Osteopontin/OPN by Western Blot. Western blot shows lysates of human adipose (diabetes) tissue. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14331) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Osteopontin/OPN at approximately 60 to 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Detection of Human Osteopontin/OPN by Simple WesternTM. Simple Western lane view shows lysates of human adipose tissue and human adipose (diabetes) tissue, loaded at 0.2 mg/mL. A specific band was detected for Osteopontin/OPN at approximately 63 kDa (as indicated) using 100 µg/mL of Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14331). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.*Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
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Detection of Osteopontin/OPN in U937 cells by Flow Cytometry U937 cells were stained with Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # mab14331, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Osteopontin/OPN
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 v beta 1, alpha v beta 3, alpha v beta 5, and non-RGD-mediated interaction with CD44 and the integrins alpha 8 beta 1 or alpha 9 beta 1. 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).
- Ann. N.Y. Acad. Sci. (1995) 760, Apr. 21.
- Senger, D.R. et al. (1996) Biochim. Biophys. Acta. 1314:13.
- Agnihotri, R. et al. (2001) J. Biol. Chem. 276:28261.
Product Datasheets
Citations for Human Osteopontin/OPN Antibody
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.
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Citations: Showing 1 - 8
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Calciprotein particle inhibition explains magnesium-mediated protection against vascular calcification
Authors: Anique D ter Braake, Coby Eelderink, Lara W Zeper, Andreas Pasch, Stephan J L Bakker, Martin H de Borst et al.
Nephrology Dialysis Transplantation
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An immunosuppressive vascular niche drives macrophage polarization and immunotherapy resistance in glioblastoma
Authors: Yang, F;Akhtar, MN;Zhang, D;El-Mayta, R;Shin, J;Dorsey, JF;Zhang, L;Xu, X;Guo, W;Bagley, SJ;Fuchs, SY;Koumenis, C;Lathia, JD;Mitchell, MJ;Gong, Y;Fan, Y;
Science advances
Applications: Western Blot -
Correlation of OPN gene expression with proliferation and apoptosis of ovarian cancer cells and prognosis of patients
Authors: H Hu, Z Liu, C Liu
Oncol Lett, 2019-01-07;17(3):2788-2794.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Osteopontin regulates HCV replication and assembly via interaction with HCV proteins and lipid droplets and by binding to receptors, αVβ3 and CD44
Authors: J Iqbal, M Sarkar-Dut, S McRae, A Ramachandr, B Kumar, G Waris
J. Virol., 2018-06-13;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Immunoprecipitation -
Thrombin-cleaved fragments of osteopontin are overexpressed in malignant glial tumors and provide a molecular niche with survival advantage.
Authors: Yamaguchi Y, Shao Z, Sharif S, Du X, Myles T, Merchant M, Harsh G, Glantz M, Recht L, Morser J, Leung L
J Biol Chem, 2012-11-30;288(5):3097-111.
Species: Human
Sample Types: Tissue Homogenates
Applications: ELISA Development -
Regional immunity in melanoma: immunosuppressive changes precede nodal metastasis.
Authors: Mansfield AS, Holtan SG, Grotz TE
Mod. Pathol., 2010-12-10;24(4):487-94.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Osteopontin induces AP-1-mediated secretion of urokinase-type plasminogen activator through c-Src-dependent epidermal growth factor receptor transactivation in breast cancer cells.
Authors: Das R, Mahabeleshwar GH, Kundu GC
J. Biol. Chem., 2004-01-02;279(12):11051-64.
Species: Human
Sample Types: Milk
Applications: Western Blot -
Abnormalities in the Motor Unit of a Fast-Twitch Lower Limb Skeletal Muscle in Huntington's Disease
Authors: Priscila Aparecida Costa Valadão, Bárbara Campos de Aragão, Jéssica Neves Andrade, Matheus Proença S. Magalhães-Gomes, Giselle Foureaux, Julliane Vasconcelos Joviano-Santos et al.
ASN Neuro
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