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Human Osteopontin/OPN Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB14334

Recombinant Monoclonal Antibody.
R&D Systems, part of Bio-Techne
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MAB14334-100
MAB14334-SP

Key Product Details

Species Reactivity

Human

Applications

Immunocytochemistry, Immunohistochemistry, Neutralization

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2579A

Product Specifications

Immunogen

Synthetic peptide containing human Osteopontin/OPN
Accession # NP_000573.1

Specificity

Detects human Osteopontin/OPN in direct ELISAs.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human Osteopontin/OPN Antibody

Osteopontin/OPN antibody in HepG2 Human Cell Line by Immunocytochemistry (ICC).

Osteopontin/OPN in HepG2 Human Cell Line.

Osteopontin/OPN was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line (positive) and Daudi human Burkitt's lymphoma cell line (negative control) using Rabbit Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14334) at 3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Osteopontin/OPN antibody in Human Kidney by Immunohistochemistry (IHC-P).

Osteopontin/OPN in Human Kidney.

Osteopontin/OPN was detected in immersion fixed paraffin-embedded sections of human kidney using Rabbit Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14334) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in convoluted tubules. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Cell Adhesion Mediated by Osteopontin/OPN and Neutralization by Human Osteopontin/OPN Antibody.

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 Rabbit Anti-Human Osteopontin/OPN Monoclonal Antibody (Catalog # MAB14334). The ND50 is typically 0.5-1.5 µg/mL.

Applications for Human Osteopontin/OPN Antibody

Application
Recommended Usage

Immunocytochemistry

3-25 µg/mL
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line

Immunohistochemistry

3-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human kidney

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 0.5-1.5 µg/mL in the presence of 1 µg/mL Recombinant Human Osteopontin/OPN.

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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 alphav beta1, alphav beta3, alphav beta5, and non-RGD-mediated interaction with CD44 and the integrins alpha8 beta1 or alpha9 beta1. 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]

Alternate Names

Eta-1, OPN, Spp1

Entrez Gene IDs

6696 (Human); 20750 (Mouse); 25353 (Rat); 281499 (Bovine)

Gene Symbol

SPP1

UniProt

Additional Osteopontin/OPN Products

Product Documents for Human Osteopontin/OPN Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Osteopontin/OPN Antibody

For research use only

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