Human Osteopontin/OPN Biotinylated Antibody

Catalog # Availability Size / Price Qty
BAF1433
Osteopontin/OPN in Human NCI‑H460 Cell Line.
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Citations (3)
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Human Osteopontin/OPN Biotinylated Antibody Summary

Species Reactivity
Human
Specificity
Detects human Osteopontin/OPN in ELISAs and Western blots. In sandwich ELISAs, less than 0.2% cross-reactivity with recombinant mouse (rm) Osteopontin and bovine Osteopontin 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 BSA as a carrier protein.
Label
Biotin

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human Osteopontin/OPN (Catalog # 1433-OP)
Immunocytochemistry
1.7-15 µg/mL
Immersion fixed NCI-H460 human large cell lung carcinoma cell line

Human Osteopontin/OPN Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Detection (Matched Antibody Pair)
0.1-0.4 µg/mL 

Use in combination with:

Capture Reagent: Human Osteopontin/OPN Antibody (Catalog # MAB14332)

Standard: Recombinant Human Osteopontin (OPN) Protein (Catalog # 1433-OP)

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

Immunocytochemistry View Larger

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.

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.
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 Biotinylated 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.

3 Citations: Showing 1 - 3
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  1. 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.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  2. 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.
    Species: Human
    Sample Types: Serum
    Applications: Multiplex Detection
  3. 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.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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