Human SPARC/Osteonectin recombinant Protein
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Human Secreted Protein Acidic and Rich in Cysteine, ON, BM-40, Basement-membrane protein-40, recombinant
Description of Human SPARC/Osteonectin
Recombinant human SPARC/Osteonectin is a single, non-glycosylated polypeptide chain containing 286 amino acids with a molecular mass of 32,7 kDa. SPARC founded a family of secreted matricellular proteins with similar domain structure. The 286 amino acid (aa), 43 kDa protein contains an N-terminal acidic region that binds calcium, a follistatin domain that contains Kazal-like sequences, and a C-terminal extracellular calcium (EC) binding domain with two EF-hand motifs. Crystal structure modeling shows that residues implicated in cell binding, inhibition of cell spreading, and disassembly of focal adhesions cluster on one face of SPARC, while a collagen binding epitope and an N-glycosylation site are opposite this face. SPARC is produced by a variety of cells e.g. fibroblasts, capillary endothelial cells, platelets and macrophages, especially in areas of tissue morphogenesis and remodeling. SPARC shows context-specific effects, but generally inhibits adhesion, spreading and proliferation, and promotes collagen matrix formation. For endothelial cells, SPARC disrupts focal adhesions and binds and sequesters PDGF and VEGF.It is abundantly expressed in bone, where it promotes osteoblast differentiation and inhibits adipogenesis. SPARC is potentially cleaved by metalloproteinases, producing an angiogenic peptide that includes the copper-binding sequence KGHK. Paradoxically, SPARC is highly expressed in many tumor types undergoing an endothelial to mesenchymal transition; its expression, however, mainly decreases the likelihood of metastasis and confers sensitivity to chemotherapy and radiation. Mature, secreted human SPARC is evolutionary conserved and shows the following aa identities: murine and rat 92%, canine 97%, bovine 99%, porcine 96% and chick 85%.
The ED50 of ≤ 3µg/ml was measured determined by its ability to inhibit the cell growth of Mv1Lu mink Lung epithelial cells. That corresponds to a specific activity of ≥ 330 u/mg.
We recommend a quick spin followed by reconstitution in sterile Water at a concentration of at least 0.1 mg/ml, which can then be further diluted into other aqueous or buffered solutions.
The lyophilized protein is stable for at least 1 year from date of receipt if stored at -20°C to –80°C. Reconstituted SPARC / Osteonectin should be stored at 4°C if entire vial will be used within 1 week. It is stable for least 3 months when stored in working aliquots below -20°C. Please avoid repeated freeze-thaw cycles.
Amino Acid Sequence
APQQEALPDE TEVVEETVAE VTEVSVGANP VQVEVGEFDD GAEETEEEVV AENPCQNHHC KHGKVCELDE NNTPMCVCQD PTSCPAPIGE FEKVCSNDNK TFDSSCHFFA TKCTLEGTKK GHKLHLDYIG PCKYIPPCLD SELTEFPLRM RDWLKNVLVT LYERDEDNNL LTEKQKLRVK KIHENEKRLE AGDHPVELLA RDFEKNYNMY IFPVHWQFGQ LDQHPIDGYL SHTELAPLRA PLIPMEHCTT RFFETCDLDN DKYIALDEWA GCFGIKQKDI DKDLVI
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