Human Osteopontin/OPN Antibody MAB14331-100: R&D Systems

Human Osteopontin/OPN Antibody

(3 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Osteopontin in direct ELISAs and Western blots. In direct ELISAs and Western blots, this antibody does not cross‑react with recombinant mouse Osteopontin.
  • Source
    Monoclonal Mouse IgG2A Clone # 223126
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Osteopontin
    Ile17-Asn300
    Accession # NP_000573.1
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Simple Western
    100 µg/mL
    See below
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.
Data Examples
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.
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.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 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).

  • References:
    1. Ann. N.Y. Acad. Sci. (1995) 760, Apr. 21.
    2. Senger, D.R. et al. (1996) Biochim. Biophys. Acta. 1314:13.
    3. Agnihotri, R. et al. (2001) J. Biol. Chem. 276:28261.
  • Long Name:
    Secreted Phosphoprotein 1 [BNSP]
  • Entrez Gene IDs:
    6696 (Human); 20750 (Mouse); 25353 (Rat); 281499 (Bovine)
  • Alternate Names:
    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
Related Research Areas
Citations:

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.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. 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, 2013;288(5):3097-111.
    Species: Human
    Sample Type: Tissue Homogenates
    Application: ELISA Development
  2. Regional immunity in melanoma: immunosuppressive changes precede nodal metastasis.
    Authors: Mansfield AS, Holtan SG, Grotz TE
    Mod. Pathol., 2011;24(4):487-94.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. 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;279(12):11051-64.
    Species: Human
    Sample Type: Milk
    Application: WB
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG2A Isotype Control

Ctrl MAB003 32  
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Mouse IgG2A Isotype Control

Ctrl MAB0031 7  
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Mouse IgG2A Isotype Control

Ctrl MAB003R
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Secondary Antibodies
Description Application Cat# Citations Images  

Mouse IgG HRP-conjugated Antibody

WB HAF007 26  
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Mouse F(ab)2 IgG (H+L) PE-conjugated Antibody

Flow F0102B 10  
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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

Flow, IHC, ICC NL007 9
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Donkey Anti-Mouse IgG Biotinylated Antibody

WB BAF018 1
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Mouse IgG HRP-conjugated Antibody

WB HAF018 9  
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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse IgG Antibody

WB AF007 4
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rat Anti-Mouse IgG2A PE-conjugated Antibody

Flow F0129 1  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Mouse IgG2A Antibody

Flow MAB0032  
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Rat Anti-Mouse IgG2A APC-conjugated Antibody

Flow F0130  
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Rat Anti-Mouse IgG2A PerCP-conjugated Antibody

Flow F0131  
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