Human Osteopontin/OPN Antibody

(9 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Osteopontin/OPN in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant rat Osteopontin, bovine Osteopontin and recombinant mouse OPN is observed.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Human milk-derived Osteopontin/OPN
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human Osteopontin/OPN (Catalog # 1433-OP)
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • 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.
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
Immunohistochemistry
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.
Immunohistochemistry
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.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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.

9 Citations: Showing 1 - 9
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Species
Applications
Sample Type
  1. Inhibition of Cellular Adhesion by Immunological Targeting of Osteopontin Neoepitopes Generated through Matrix Metalloproteinase and Thrombin Cleavage.
    Authors: Jurets A, Le Bras M, Staffler G, Stein G, Leitner L, Neuhofer A, Tardelli M, Turkof E, Zeyda M, Stulnig T
    PLoS ONE, 2016;11(2):e0148333.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Osteopontin promotes the invasive growth of melanoma cells by activating integrin alphavbeta3 and down-regulating tetraspanin CD9.
    Authors: Yin M, Soikkeli J, Jahkola T, Virolainen S, Saksela O, Holtta E
    Am J Pathol, 2014;184(3):842-58.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  3. Cell stress induces upregulation of osteopontin via the ERK pathway in type II alveolar epithelial cells.
    Authors: Kato, Aki, Okura, Takafumi, Hamada, Chizuru, Miyoshi, Seigo, Katayama, Hitoshi, Higaki, Jitsuo, Ito, Ryoji
    PLoS ONE, 2014;9(6):e100106.
    Species: Human
    Sample Type: Whole Cells
    Application: Neut
  4. CD44v6 is a marker of constitutive and reprogrammed cancer stem cells driving colon cancer metastasis.
    Authors: Todaro M, Gaggianesi M, Catalano V, Benfante A, Iovino F, Biffoni M, Apuzzo T, Sperduti I, Volpe S, Cocorullo G, Gulotta G, Dieli F, De Maria R, Stassi G
    Cell Stem Cell, 2014;14(3):342-56.
    Species: Human
    Sample Type: Whole Cells
    Application: Functional Assay
  5. Mechanism of hepatitis C virus (HCV)-induced osteopontin and its role in epithelial to mesenchymal transition of hepatocytes.
    Authors: Iqbal J, McRae S, Banaudha K, Mai T, Waris G
    J Biol Chem, 2013;288(52):36994-7009.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Regulation and function of immunosuppressive molecule human leukocyte antigen G5 in human bone tissue.
    Authors: Deschaseaux F, Gaillard J, Langonne A, Chauveau C, Naji A, Bouacida A, Rosset P, Heymann D, De Pinieux G, Rouas-Freiss N, Sensebe L
    FASEB J, 2013;27(8):2977-87.
    Species: Human
    Sample Type: Whole Cells
    Application: IF
  7. Osteopontin is a prognostic factor for survival of acute myeloid leukemia patients.
    Authors: Liersch R, Gerss J, Schliemann C, Bayer M, Schwoppe C, Biermann C, Appelmann I, Kessler T, Lowenberg B, Buchner T, Hiddemann W, Muller-Tidow C, Berdel WE, Mesters R
    Blood, 2012;119(22):5215-20.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  8. Secreted osteopontin is highly polymerized in human airways and fragmented in asthmatic airway secretions.
    Authors: Arjomandi M, Frelinger J, Donde A
    PLoS ONE, 2011;6(10):e25678.
    Species: Human
    Sample Type: Recombinant Protein
    Application: WB
  9. The crucial role of cyclooxygenase-2 in osteopontin-induced protein kinase C alpha/c-Src/IkappaB kinase alpha/beta-dependent prostate tumor progression and angiogenesis.
    Authors: Jain S, Chakraborty G, Kundu GC
    Cancer Res., 2006;66(13):6638-48.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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