Human Osteopontin/OPN Antibody

Catalog # Availability Size / Price Qty
MAB14331-SP
MAB14331-500
MAB14331-100
Detection of Human Osteopontin/OPN by Western Blot.
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Product Details
Citations (5)
FAQs
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Human Osteopontin/OPN Antibody Summary

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 either lyophilized or as a 0.2 µm filtered solution in PBS.

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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.

Scientific Data

Western Blot Detection of Human Osteopontin/OPN antibody by Western Blot. View Larger

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.

Simple Western Detection of Human Osteopontin/OPN antibody by Simple Western<SUP>TM</SUP>. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 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

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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.

5 Citations: Showing 1 - 5
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  1. 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;17(3):2788-2794.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. 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;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Immunoprecipitation
  3. 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 Types: Tissue Homogenates
    Applications: ELISA Development
  4. 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 Types: Whole Tissue
    Applications: IHC-P
  5. 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 Types: Milk
    Applications: Western Blot

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Isotype Controls

Mouse IgG2A Isotype Control

Reconstitution Buffers

Secondary Antibodies

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