Active Osteopontin (OPN)

骨桥素(OPN)活性蛋白

[ PROPERTIES ]

Source: eukaryotic expression. 

Host: 293 cell

Residues: Ile17~Asn287

Tags: N-terminal His-tag

Purity: >95%

Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 1mM EDTA, 1mM DTT, 0.01% sarcosyl, 5% trehalose, and Proclin300. Applications: Cell culture; Activity Assays; In vivo assays. 

(May be suitable for use in other assays to be determined by the end user.)

Predicted isoelectric point: 4.3

Predicted Molecular Mass: 32.2kDa

Accurate Molecular Mass: 60kDa as determined by SDS-PAGE reducing conditions. 

Phenomenon explanation:

The possible reasons that the actual band size differs from the predicted are as follows:

1. Splice variants: Alternative splicing may create different sized proteins from the same gene. 2. Relative charge: The composition of amino acids may affects the charge of the protein. 3. Post-translational modification: Phosphorylation, glycosylation, methylation etc. 4. Post-translation cleavage: Many proteins are synthesized as pro-proteins, and then cleaved to

give the active form. 5. Polymerization of the target protein: Dimerization, multimerization etc.

[ USAGE ]

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0mg/mL. Do not vortex. 

[ STORAGE AND STABILITY ]

Storage: Avoid repeated freeze/thaw cycles. 

Store at 2-8℃ for one month. 

Aliquot and store at -80℃ for 12 months. 

Stability Test: The thermal stability is described by the loss rate. The loss ratewas determined by accelerated thermal degradation test, that is, incubate the protein at 37℃ 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. 

[ SEQUENCE ]

[ ACTIVITY ]

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-kappa B pathways in MCF-7 breast cancer cell line. Briefly, MCF-7 cells were seeded overnight at a density of 1x105 cells/mL, and treated with or without 200ng/mL OPN for 24h and MCP-1 levels in the cell supernatant were determined by ELISA. Result: MCP-1 levels in the cell supernatant of MCF-7 cells increased significantly after stimulated with OPN, the data was shown in Table 1 and Figure 1.