Buffer Formulation	20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose and Proclin300.
Traits	Freeze-dried powder
Purity	> 95%
Isoelectric Point	4.3
Applications	Cell culture; Activity Assays.

ACTIVITY TEST

Osteopontin (OPN), a multifunctional phosphorylated glycoprotein, plays an important role in neutrophil recruitment and was found to induce the expression of proinflammatory chemokines including MCP-1 and MIP-1β which promote migration and recruitment of inflammatory cells. It has been reported that OPN induces MCP-1 expression through the NF-kappaB pathways in MCF-7 breast cancer cell line. To test the bioactivity of recombinant rat OPN, MCF-7 cells were seeded into 24-well plate at a density of 1x105 cells/mL , and allowed to attach overnight before treated with certain concentrations of recombinant rat OPN for 48h and MCP-1 levels in the cell supernatant were determined by ELISA (SEA087Hu). MCP-1 levels in the cell supernatant of MCF-7 cells increased significantly after stimulated with OPN which was shown in Figure1, the EC50 was 9.83 ug/ml.

 

USAGE

 

Reconstitute in 20mM Tris, 150mM NaCl (PH8.0) 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.

 

Image


SDS-PAGE Image

 

Figure. Western Blot; Sample: Recombinant OPN, Rat.

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