Human Osteopontin/OPN Antibody
R&D Systems, part of Bio-Techne | Catalog # AF1433
Key Product Details
Species Reactivity
Validated:
Cited:
Applications
Validated:
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Label
Antibody Source
Product Specifications
Immunogen
Specificity
Clonality
Host
Isotype
Endotoxin Level
Scientific Data Images for Human Osteopontin/OPN Antibody
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.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.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.Applications for Human Osteopontin/OPN Antibody
Dual RNAscope ISH-IHC Compatible
Sample: Immersion fixed paraffin-embedded sections of human kidney
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.
Immunocytochemistry
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line
Immunohistochemistry
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue
Neutralization
Formulation, Preparation, and Storage
Purification
Reconstitution
Formulation
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Stability & Storage
- 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
- Ann. N.Y. Acad. Sci. (1995) 760, Apr. 21.
- Senger, D.R. et al. (1996) Biochim. Biophys. Acta. 1314:13.
- Agnihotri, R. et al. (2001) J. Biol. Chem. 276:28261.
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Additional Osteopontin/OPN Products
Product Documents for Human Osteopontin/OPN Antibody
Product Specific Notices for Human Osteopontin/OPN Antibody
For research use only