Invitrogen™ Human VEGF-165, Recombinant Protein

Description

VEGF-165 (human) Recombinant Protein, RVEGFI, from Invitrogen.

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. Recombinant Human VEGF produced in E.Coli is a double, non-glycosylated, polypeptide chain containing 165 amino acids and having a molecular mass of 38231 Dalton. Mature human VEGF165 was expressed in Pichia pastoris, KM71. GenBank Accession # 003376, amino acids 27 - 191. For protein verification, the expressed protein was N-terminal sequenced and begins with the following 10 amino acids: EGGGQNHHEV. Purity is >95% by SDS-PAGE analyses. Activity: The biological activity of recombinant human VEGF165 was determined by measuring the proliferation of human umbilical vein endothelial cells (HUVEC). Use: For in vitro applications. A dose-response assay should be performed to determine the optimal concentration for specific experiments. Reconstitution and Dilution: Store lyophilized material at 4°C. Reconstitute in 1 ml of distilled or deionized water. Dilute in a buffer-containing carrier protein, aliquot, and store at or below -70°C, avoiding multiple freeze-thaw cycles. PBS with 1% BSA or a neutral media containing <10% FCS are suitable buffers.