Recombinant Mouse Osteopontin/OPN Protein

R&D Systems, part of Bio-Techne | Catalog # 441-OP

R&D Systems, part of Bio-Techne
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Carrier Free
441-OP-050/CF
441-OP-200/CF
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With Carrier
441-OP-050
441-OP-200
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Key Product Details

Source

NS0

Accession #

Q547B5

Conjugate

Unconjugated

Applications

Bioactivity

View all Osteopontin/OPN Proteins and Enzymes »

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Osteopontin/OPN protein
Leu17-Asn294 (Glu99Gly), with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu17

Predicted Molecular Mass

31.5 kDa

SDS-PAGE

30 kDa and a 65 kDa doublet, reducing conditions

Activity

Measured by the ability of the immobilized protein to support the adhesion of HEK293 human embryonic kidney cells. Agnihotri, R. et al. (2001) J. Biol. Chem. 276:28261.
When 1 x 105 cells/well are added to a Recombinant Mouse (rm) Osteopontin/OPN coated plate, cell adhesion is enhanced in a dose-dependent manner after 1 hour incubation at 37 °C. The ED50  for this effect is 0.25-1.5 µg/mL. Recombinant Human Coagulation Factor II/Thrombin (Catalog # 1473-SE) proteolytic treatment of this rmOsteopontin can increase HEK293 cell adhesion by about 10-fold.

Reviewed Applications

Read 1 review rated 5 using 441-OP in the following applications:

Formulation, Preparation and Storage

Carrier Free
What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

Carrier: 441-OP
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Carrier Free: 441-OP/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 200 μg/mL in sterile PBS.
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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Osteopontin/OPN

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).

References

  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.

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

Q547B5

Additional Osteopontin/OPN Products

Product Documents for Recombinant Mouse Osteopontin/OPN Protein

Product Datasheets

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Recombinant Mouse Osteopontin/OPN Protein

For research use only

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