Human Osteopontin/OPN Antibody

Catalog # Availability Size / Price Qty
AF1433
AF1433-SP
Human Osteopontin/OPN Antibody in Immunocytochemistry
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Citations (16)
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Human Osteopontin/OPN Antibody Summary

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 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
ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14332R).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human Osteopontin (OPN) DuoSet ELISA Kit (Catalog # DY1433) for convenient development of a sandwich ELISA or the Human Osteopontin (OPN) Quantikine ELISA Kit (Catalog # DOST00) for a complete optimized ELISA.

 
Immunohistochemistry
5-15 µg/mL
See below
Immunocytochemistry
1-15 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

Immunocytochemistry Osteopontin/OPN  in HepG2 Human Cell Line. View Larger

Osteopontin/OPN in HepG2 Human Cell Line. Osteopontin/OPN was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line (positive staining; left panel) and Daudi human Burkitt's lymphoma cell line (negative staining; right panel) using Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) at 1.7 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 Cells on Coverslips.

Immunohistochemistry Osteopontin/OPN in Human Breast Cancer Tissue. View Larger

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. View Larger

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.

Neutralization Cell Adhesion Mediated by Osteopontin/OPN and Neutralization by Human Osteopontin/OPN Antibody. View Larger

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.

Human Osteopontin/OPN ELISA Standard Curve. Recombinant Human Osteopontin/OPN protein was serially diluted 2-fold and captured by Mouse Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14332R) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Osteopontin/OPN Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1433) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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.

16 Citations: Showing 1 - 10
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  1. Differential expression patterns of Toll Like Receptors and Interleukin-37 between calcific aortic and mitral valve cusps in humans
    Authors: A Kapelouzou, C Kontogiann, DI Tsilimigra, G Georgiopou, L Kaklamanis, L Tsourelis, DV Cokkinos
    Cytokine, 2019;116(0):150-160.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC
  2. Osteopontin contributes to effective neutrophils recruitment, IL-1? production, and apoptosis in Aspergillus fumigatus keratitis
    Authors: G Zhao, M Hu, C Li, J Lee, K Yuan, G Zhu, C Che
    Immunol. Cell Biol., 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  3. Phenotypic and Functional Characterization of Peripheral Sensory Neurons derived from Human Embryonic Stem Cells
    Authors: AJ Alshawaf, S Viventi, W Qiu, G D'Abaco, B Nayagam, M Erlichster, G Chana, I Everall, J Ivanusic, E Skafidas, M Dottori
    Sci Rep, 2018;8(1):603.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  4. Osteopontin mediates glioblastoma-associated macrophage infiltration and is a therapeutic target
    Authors: J Wei, A Marisetty, B Schrand, K Gabrusiewi, Y Hashimoto, M Ott, Z Grami, LY Kong, X Ling, HG Caruso, S Zhou, YA Wang, GN Fuller, JT Huse, E Gilboa, N Kang, X Huang, R Verhaak, S Li, AB Heimberger
    J. Clin. Invest., 2018;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  5. Single-Cell RNA-Seq Reveals the Transcriptional Landscape and Heterogeneity of Aortic Macrophages in Murine Atherosclerosis
    Authors: C Cochain, E Vafadarnej, P Arampatzi, P Jaroslav, H Winkels, K Ley, D Wolf, AE Saliba, A Zernecke
    Circ. Res., 2018;0(0):.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  6. Osteopontin - Metallothionen I/II Interactions In Experimental Autoimmunune Encephalomyelitis
    Authors: H Jakovac, TG Kezele, S Šu?urovi?, B Mulac-Jeri, B Radoševi?-
    Neuroscience, 2017;0(0):.
    Species: Rat
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  7. Generation of iPSC-derived limb progenitor-like cells for stimulating phalange regeneration in the adult mouse
    Authors: Y Chen, H Xu, G Lin
    Cell Discov, 2017;3(0):17046.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  8. 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 Types: Cell Lysates
    Applications: Western Blot
  9. 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 Types: Whole Cells
    Applications: Functional Assay
  10. 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 Types: Whole Cells
    Applications: Neutralization
  11. 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 Types: Whole Cells
    Applications: Neutralization
  12. 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 Types: Cell Lysates
    Applications: Western Blot
  13. 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 Types: Whole Cells
    Applications: IHC
  14. 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 Types: Whole Tissue
    Applications: IHC-P
  15. 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 Types: Recombinant Protein
    Applications: Western Blot
  16. 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 Types: Cell Lysates
    Applications: Western Blot

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