Sialoprotein, Bone, Recombinant, Human (BSP)

IBSP (integrin-binding sialoprotein; also BSP or bone sialoprotein (II)) is a 55-75 kD, secreted, variably glycosylated, monomeric noncollagenous member of the SIBLING family of extracellular matrix (ECM) proteins (1-3). It is principally associated with the early stages of bone mineralization. BSP is synthesized as a 317 amino acid (aa) precursor that contains a 16 aa signal sequence and a 301 aa mature region (4-6). BSP is highly glycosylated, sulfated, and phosphorylated. Phosphorylation may impact HAp growth, while carbohydrate may regulate cell adhesion (1, 3, 13). Mature human BSP is 70%, 72%, 78%, and 72% aa identical to porcine, rat, canine, and mouse BSP, respectively. BSP is synthesized by megakaryocytes/ platelets, osteoblasts, osteocytes, odontoblasts, osteoclasts, and bone marrow stromal cells (14-17).\n\nDescription: \nRecombinant human BSP is composed of a Human CD33 Signal Peptide (Met 1-Arg 399), followed by Human IBSP (Phe 17-Gln 317) (Accession #AAC95490), and with a poly-his tag at the C-terminus. \n\nMolecular Mass: \nRecombinant human BSP migrates as an ~75-90kD protein in SDS-PAGE under reducing conditions.\n\n\nActivity: \nMeasured by the ability of the immobilized protein to support the adhesion of MCF-7 cells. When 5x10e4 cells/well are added to recombinant human IBSP-coated plates (3ug/ml with 100ul/well), ~60-80% will adhere after 30 minutes at 37 degrees C. Optimal concentration depends on cell type as well as the application or research objectives.\n\nStorage and Stability: Lyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Reconstituted product is stable for 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

S1013-34L

Size

50ug

Form

Supplied as a lyophilized powder in 20mM MES, 0.5M sodium chloride, pH 6.5. Reconstitute with sterile PBS, 0.1% HSA or BSA to (same/more than)100ug/ml.

Purity

~90%, as determined by SDS-PAGE and visualized by silver stain. Endotoxin: (same/less than)1.0EU/ug (LAL).

References

1. Qin, C. et al. (2004) Crit. Rev. Oral Biol. Med. 15:126. 2. Alford, A.I. and K.D. Hankenson (2006) Bone 38:749. 3. Ganss, B. et al. (1999) Crit. Rev. Oral Biol. Med. 10:79. 4. Fisher, L.W. et al. (1990) J. Biol. Chem. 265:2347. 5. Kerr, J.M. et al. (1993) Genomics 17:408. 6. Kim, R.H. et al. (1994) Matrix Biol. 14:31. 7. Zaia, J. et al. (2001) Biochemistry 40:12983. 8. Tye, C.E. et al. (2005) J. Biol. Chem. 280:13487. 9. Stubbs, J.T. et al. (1997) J. Bone Miner. Res. 12:1210. 10. Tye, C.E. et al. (2003) J. Biol. Chem. 278:7949. 11. Miyauchi, A. et al. (1991) J. Biol. Chem. 266:20369. 12. Hakki, S.S. et al. (2006) J. Periodontol. 77:167. 13. Wazen, R.M. et al. (2007) J. Histochem. Cytochem. 55:35. 14. Kacena, M.A. et al. (2006) Bone 39:978. 15. Bianco, P. et al. (1991) Calcif. Tissue Int. 49:421. 16. Chen, J. et al. (1992) J. Bone Miner. Res. 7:987. 17. Kreke, M.R. et al. (2005) Bone 36:1047.