Human Osteopontin/OPN Biotinylated Antibody Summary
Human Osteopontin/OPN Sandwich Immunoassay
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.
Osteopontin/OPN in Human NCI‑H460 Cell Line. Osteopontin/OPN was detected in immersion fixed NCI‑H460 human large cell lung carcinoma cell line using Goat Anti-Human Osteopontin/OPN Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1433) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and 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.
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).
- 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.
Citations for Human Osteopontin/OPN Biotinylated Antibody
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Bone marrow adipose tissue does not express UCP1 during development or adrenergic-induced remodeling
Authors: CS Craft, H Robles, MR Lorenz, ED Hilker, KL Magee, TL Andersen, WP Cawthorn, OA MacDougald, CA Harris, EL Scheller
Sci Rep, 2019;9(1):17427.
Sample Types: Whole Tissue
A multiplex immunoassay of serum biomarkers for the detection of uveal melanoma
Authors: J Song, SL Merbs, LJ Sokoll, DW Chan, Z Zhang
Clin Proteomics, 2019;16(0):10.
Sample Types: Serum
Applications: Multiplex Detection
Gene expression signatures for tumor progression, tumor subtype, and tumor thickness in laser-microdissected melanoma tissues.
Authors: Jaeger J, Koczan D, Thiesen HJ, Ibrahim SM, Gross G, Spang R, Kunz M
Clin. Cancer Res., 2007;13(3):806-15.
Sample Types: Cell Lysates
Applications: Western Blot
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