Human Vascular Endothelial Growth Factor (121), Sf9 (Recombinant)

Recombinant Human Vascular Endothelial Growth Factor-121 glycosylated (VEGF-121 )

Source: Sf9, Insect Cells

Background: Vascular endothelial growth factor-A was originally isolated from tumor cells and referred to as Tumor Angiogenesis Factor or Vascular Permeability Factor. Although expressed at high levels in certain tumor-derived cells it is produced by a wide variety of cell types. In addition to stimulating vascular growth and vascular permeability it may play a role in stimulating Vasolidation via nitric oxide-dependent pathways. Alternative splicing of the mRNA for VEGF-A results in several isoforms of the protein being produced. Rat and bovine VEGF are one amino acid shorter than the human factor, and the bovine and human sequences show a homology of 95 percent. In contrast to other factors mitogenic for endothelial cells such as FGF-1 , FGF-2 and PDGF, VEGF is synthesized as a precursor containing a typical hydrophobic secretory signal sequence of 26 amino acids. Glycosylation is not required for efficient secretion of VEGF.

Description: Recombinant Human VEGF-121 produced in insect cells is a double, non-glycosylated, polypeptide chain containing 121 amino acids and having a molecular mass of 28423 Dalton. VEGF121 circulates more freely than other VEGF forms, which bind more tightly with vascular heparin sulfates. Recombinant Human VEGF-121 is purified by proprietary chromatographic techniques.

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

Formulation: The protein was lyophilized from a concentrated (1mg/ml) solution with no additives.

Solubility: It is recommended to reconstitute the lyophilized Human VEGF-121 in sterile 18MO-cm H2O not less than 100 g/ml, which can then be further diluted to other aqueous solutions.

Stability: Lyophilized Recombinant VEGF-121 although stable at room temperature for 3 weeks, should be stored desiccated below -180C. Upon reconstitution VEGF-121 should be stored at 40C between 2-7 days and for future use below -180C. For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA). 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.

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

Biological Activity: Recombinant Human VEGF-121 is fully biologically active when compared to standards. Determined by the dose-dependent stimulation of the proliferation of human umbilical vein endothelial cells (HUVEC) using a concentration range of 0.2-0.4 ng/ml, corresponding to a Specific Activity of 5 x 105 IU/mg.

Endotoxin: Less than 0.1 ng/ g (IEU/ g) of VEGF 121.

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-121 as a Reference Standard.

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