VEGF 165

VEGF (Vascular endothelial growth factor) is the only growth factor that stimulates vascular permeability. It promotes endothelial proliferation and survival, angiogenesis, vasculognesis and inhibits apoptosis. VEGF mitogenic activity is specific for endothelial cell and that makes it distinct among other growth factors. VEGF is thought to be important in the pathophysiology of neuronal and other tumors, by functioning as a prompter of angiogenesis for human gliomas. Human VEGF occurs in several molecular variants and the 165 form is the most common form in most tissues. Biological activities of VEG F are not species specific and glycosylation is not required for biological activity.

Produced in the endosperm tissue of barley grain (Hordeum vulgare).

PBS pH 7.2, sterile filtered through 0.2 µm filter.

Greater than 95% by SDS-PAGE gel analysis.

Note: Always centrifuge the vial before opening. It is recommended to reconstitute the lyophilized protein in sterile buffered saline to a concentration of no less than 100 µg/ml. For long term storage of the reconstituted solution it is recommended to add a carrier protein (0.1% HSA or BSA).

The lyophilized protein, though stable at room temperature for few weeks, is best stored at -20°C. Reconstituted protein should be used immediately or stored in working aliquots at -20°C. Avoid repeated freeze-thaw cycles.

The bioactivity of ISOkine® recombinant human VEGF 165 was assayed by measuring its dose dependent effect on proliferation of Human Umbilical Vein Endothelial Cells (HUVEC).

The ED50 for this effect is < 10 ng/ml corresponding to specific activity of > 1 x 10e5 U/mg. Optimal concentration should be determined for specific applications and cell lines.

We recommend that the optimal concentration for each specific application be determined by an initial dose-response assay. Each batch of this ISOkine® growth factors is tested for bioactivity and verified to have comparable activity to a commercial source.

Endotoxin level is less than 0.005ng per µg of ISOkine® product (0.05EU/µg).

Purified ISOkine® product carries no pyrogenic or pro-inflammatory contaminants, as assayed with monocyte activation test using Human 10-plex Cytokine Assay measuring IL-6, TNF-alpha and IL-1beta induction. ** Ref. The Blood Bank, University Hospital of Iceland, Reykjavik, Iceland

Recombinant human VEGF165 contains 165 amino acids and a 16 a.a. Histidine based tag for a total length of 181 a. a. and has a predicted molecular mass of 21.3 kDa. As a result of glycosylation, the recombinant protein migrates with an apparent molecular mass of 26-28 kDa in SDS PAGE.