Osteopontin/OPN Antibody (2139B) [Unconjugated]

Western blot shows conditioned media from Neuro-2A mouse neuroblastoma cell line (negative control) and L-929 mouse fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Mouse Osteopontin/OPN Monoclonal Antibody (Catalog # MAB808) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Osteopontin/OPN at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Osteopontin/OPN was detected in immersion fixed RAW 264.7 mouse monocyte/macrophage cell line using Rabbit Anti-Mouse Osteopontin/OPN Monoclonal Antibody (Catalog # MAB808) 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 Non-adherent Cells.

Osteopontin/OPN was detected in perfusion fixed frozen sections of mouse kidney using Rabbit Anti-Mouse Osteopontin/OPN Monoclonal Antibody (Catalog # MAB808) at 3 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to convoluted tubules. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Silencing of OPN impairs the growth of AB22 cells in vivo. (A, B)In vivo growth of 5x104 AB22 sh- scrambled or AB22 shOPN, injected intra-thoracically in BALB/c mice. (A) IVIS in vivo imaging luminescence signal, mean+/-SEM values of 5 mice for sh-scrambled, 5 for shOPN; (B) luminescence signal values of each single mouse. (C) Histological quantification of number of tumor foci, and (D) total tumor area. Data are shown as mean +/- SEM; (A, B Two-way ANOVA; C, D: One-way ANOVA; E: Unpaired t test with Welch’s correction). E, F) immunohistochemistry for OPN in explanted tumors and representative pictures, bars represent 100 µm. (G) Representative pictures of explanted lungs from mice bearing control AB22 sh-scrambled cells (left), AB22 shOPN cells at day 17 (middle) and AB22 shOPN cells at day 56 (right). Bars represent 2.5 mm. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/37398648), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, IHC, ICC/IF
• Clone: 2139B
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated

Osteopontin/OPN Antibody (2139B) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Osteopontin/OPN
  Leu17-Asn294
  Accession # Q547B5
• Specificity: Detects mouse Osteopontin /OPN in direct ELISAs.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Purity Statement: Protein A or G purified from cell culture supernatant

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 3-25 ug/mL
  • Immunohistochemistry 3-25 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Osteopontin/OPN Antibody (2139B) [Unconjugated]

• 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

Background

Osteopontin (OPN), previously called SPP1 (secreted phosphoprotein 1), Eta-1 (early T lymphocyte activation 1) or BSP (bone sialoprotein), is a secreted molecule in the SIBLING (small integrin-binding ligand N-linked glycoprotein) family of non-collagenous matricellular proteins (1-3). Mouse OPN is synthesized as a 294 amino acid (aa) precursor protein with a 16 aa signal peptide and a 278 aa mature protein (3). Mature mouse OPN shares 79% and 64% aa sequence identity with rat and human OPN, respectively. OPN is highly acidic and has 26 potential Ser/Thr phosphorylation sites and a C‑terminal CD44 binding site (1-4). Depending on tissue-specific modification by O- and N-glycosylation, sulfation, phosphorylation and transglutamination, OPN can be detected at 45-75 kDa (5, 6). The central region of OPN contains RGD and non-RGD binding sites for multiple integrins (3, 4). Adjacent to the RGD motif is the sequence SLAYGLR (SVVYGLR in human) which serves as a cryptic binding site for additional integrins: it is masked in full length OPN but is exposed following OPN cleavage by thrombin in tumors and sites of tissue injury
(6-8). OPN can also be cleaved by MMP-3, -7, -9, and -12 within the SLAYGLR motif and at sites closer to the C-terminus (8, 9). OPN is widely expressed and is prominent in mineralized tissues. It inhibits bone mineralization and kidney stone formation, and promotes inflammation and cell ad­hesion and migration (1, 2, 4, 6). Its expression is up-regulat­ed during inflammation, obesity, atherosclerosis, cancer, and tissue damage, and contributes to the pathophysiology of these conditions (1, 2, 6, 9, 10).

1. Scatena, M. et al. (2007) Arterioscler. Thromb. Vasc. Biol. 27:2302.
2. Rangaswami, H. et al. (2006) Trends Cell Biol. 16:79.
3. Miyazaki, Y. et al. (1990) J. Biol. Chem. 265:14432.
4. Weber, G.F. et al. (2002) J. Leukoc. Biol. 72:752.
5. Keykhosravani, M. et al. (2005) Biochemistry 44:6990.
6. Kazanecki, C.C. et al. (2007) J. Cell. Biochem. 102:912.
7. Senger, D.R. et al. (1994) Mol. Biol. Cell 5:565.
8. Yokosaki, Y. et al. (2005) Matrix Biol. 24:418.
9. Takafuji, V. et al. (2007) Oncogene 26:6361.
10. Kiefer, F.W. et al. (2010) Diabetes 59:935.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Osteopontin/OPN Antibody (MAB808)(5)

Publications using MAB808

• Elisabeth Digifico, Marco Erreni, Laura Mannarino, Sergio Marchini, Aldo Ummarino, Clément Anfray et al. Important functional role of the protein osteopontin in the progression of malignant pleural mesothelioma Frontiers in Immunology 2023-06-16 [PMID: 37398648] (Immunohistochemistry, Mouse)
• F Yang, S Kalantari, B Ruan, S Sun, Z Bian, JL Guan Autophagy inhibition prevents lymphatic malformation progression to lymphangiosarcoma by decreasing osteopontin and Stat3 signaling Nature Communications, 2023-02-22;14(1):978. 2023-02-22 [PMID: 36813768] (Western Blot, Mouse)
• Ashley L. Farris, Dennis Lambrechts, Yuxiao Zhou, Nicholas Y. Zhang, Naboneeta Sarkar, Megan C. Moorer et al. 3D-printed oxygen-releasing scaffolds improve bone regeneration in mice Biomaterials 1/1/2022 [PMID: 34922272]
• M Herrmann, A Babler, I Moshkova, F Gremse, F Kiessling, U Kusebauch, V Nelea, R Kramann, RL Moritz, MD McKee, W Jahnen-Dec Lumenal calcification and microvasculopathy in fetuin-A-deficient mice lead to multiple organ morbidity PLoS ONE, 2020-02-19;15(2):e0228503. 2020-02-19 [PMID: 32074120] (IHC, Mouse)
• LA Mesentier-, LC Teixeira-P, F Gubert, JF Vasques, AJ Silva-Juni, L Chimeli-Or, G Nascimento, R Mendez-Ote, MF Santiago Long-term neuronal survival, regeneration, and transient target reconnection after optic nerve crush and mesenchymal stem cell transplantation Stem Cell Res Ther, 2019-04-17;10(1):121. 2019-04-17 [PMID: 30995945] (IHC, Rat)

Bioinformatics

• Uniprot:
  • Q547B5()