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Active Osteonectin (ON)

SPARC; BM-40; Secreted Protein,Acidic,Cysteine-Rich; Basement-membrane protein 40; Secreted protein acidic and rich in cysteine

ACTIVITY TEST

SPARC, also known as osteonectin（ON）, is the founding member of a family of secreted matricellular proteins with similar domain structure. The 302 amino acid (aa), 43 kDa protein contains a 17 aa signal sequence, an N-terminal acidic region that binds calcium, a follistatin domain containing Kazal-like sequences, and a C-terminal extracellular calcium (EC) binding domain with two EF-hand motifs. SPARC is produced by 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. Besides, Collagen Type I Alpha 2 (COL1a2)) has been identified as an interactor of SPARC, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human SPARC and recombinant human COL1a2. Briefly, SPARC was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to COL1a2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-SPARC pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human SPARC and recombinant human COL1a2 was shown in Figure 1, and this effect was in a dose dependent manner.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
J Mol Med (Berl) Growth hormone replacement therapy regulates microRNA-29a and targets involved in insulin resistance PubMed: 26199111
Muscle Nerve Effect of resistance ladder training on SPARC expression in skeletal muscle of hindlimb immobilized rats PubMed: 26467437
Medicine (Baltimore) An Attempt to Evaluate Selected Aspects of “Bone–Fat Axis” Function in Healthy Individuals and Patients With Pancreatic Cancer PubMed: 26266370
British Journal of Cancer A proteomics-based approach identifies secreted protein acidic and rich in cysteine as a prognostic biomarker in malignant pleural mesothelioma Pubmed:26889976
Journal of diabetes and its complications Associations between FGF21, osteonectin and bone turnover markers in type 2 diabetic patients with albuminuria. pubmed:27916484
American journal of the medical sciences Association of Bone Metabolic Markers With Diabetic Retinopathy and Diabetic Macular Edema in Elderly Chinese Individuals With Type 2 Diabetes Mellitus pubmed:29078839
Translational Research in Oral Oncology Testican 1 (SPOCK1) and protein tyrosine phosphatase, receptor type S (PTPRS) show significant increase in saliva of tobacco users with oral cancer Doi: 10.1177/2057178X18800534
[ENGINEERING OF BIOMATERIALS Osteoconductive potential of PLGA/bioglass composite biomaterials
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