Rat Vascular Endothelial Growth Factor-C (152) (Recombinant)

Recombinant Rat Vascular Endothelial Growth Factor-C 152 a.a. (VEGF-C152)

Source: Sf9, Insect Cells

Background: VEGF-C152S is a point mutant generated by the replacement of the second conserved Cys residue of the recombinant processed VEGF-C by a Ser residue. VEGF-C 152S is analog to the human VEGF-C 156S mutant and only active toward VEGFR-3/FLT-4 but, unlike wild type VEGF-C, is unable to bind to and to activate signalling through VEGFR-2/KDR. VEGF-C152S was inactive in the vascular permeability assay and did not increase migration of the capillary endothelial cells, indicating that these VEGF-like effects of VEGF-C require VEGFR-2 binding. VEGF-C, also known as Vascular Endothelial Growth Factor Related Protein (VRP), is a recently discovered VEGF growth factor family member that is most closely related to VEGF-D. The rat VEGF-C cDNA encodes a pre-pro-protein of 416 amino acids residues. It is almost identical to the mouse VEGF-C protein. Similar to VEGF-D, VEGF-C has a VEGF homology domain spanning the middle third of the precursor molecule and long N- and C-terminal extensions. In adults, VEGF-C is highly expressed in heart, placenta, ovary and small intestine. Recombinant rat VEGF-C, lacking the N- and C-terminal extensions and containing only the middle VEGF homology domain, forms primarily non-covalently linked dimers. This protein is a ligand for both VEGFR-2/KDR and VEGFR-3/FLT -4. Since VEGFR-3 is strongly expressed in lymphatic endothelial cells, it has been postulated that VEGF-C is involved in the regulation of the growth and/or differentiation of lymphatic endothelium. Although recombinant rat VEGF-C is also a mitogen for vascular endothelial cells, it is much less potent than VEGF-A.

Description: recombinant rat VEGF-C 152 contains 152 amino acids residues and was fused to a His-tag (6x His) at the C-terminal end. As a result of glycosylation VEGF-C migrates as an 18-24 kDa protein in SDS-PAGE under reducing conditions.

Physical Appearance: Sterile Filtered White lyophilized (freeze-dried) powder.

Formulation: The protein was lyophilized from a concentrated (1mg/ml) solution with BSA.

Solubility: It is recommended to reconstitute the lyophilized Recombinant Rat VEGF-C 152 in sterile 18MO-cm H2O not less than 100 g/ml, which can then be further diluted to other aqueous solutions.

Stability: Lyophilized Rat VEGF-C152 although stable at room temperature for 3 weeks, should be stored desiccated below -180C. Upon reconstitution VEGF-C 152 should be stored at 40C between 2-7 days and for future use below -180C. Please prevent freeze-thaw cycles.

Purity: Greater than 90.0% as determined by: (a) Analysis by RP-HPLC. (b) Anion-exchange FPLC. (c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.

Amino acid sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Cys-Asp-Leu-Pro-Glu.

Dimers and aggregates: Less than 1% as determined by silver-stained SDS-PAGE gel analysis.

Biological Activity: Measured by its ability to stimulate phosphorylation of the VEGFR-3/FLT-4 receptor in porcine aortic endothelial cells (PAE/FLT -4 cells). The ED50 for this effect is typically 150-300 ng/ml, corresponding to a specific activity of 5 x 103 Units/mg.

Endotoxin: Less than 0.1 ng/ g (IEU/ g) of Rat VEGF-C152.

Protein content: Protein quantitation was carried out by two independent methods: 1. UV spectroscopy at 280 nm. 2. Analysis by RP-HPLC, using a calibrated solution of VEGF-C152 as a Reference Standard.

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