Vascular endothelial growth factor (VEGF), also known as vascular permeability factor (VPF) or vasculotropin, is a homodimeric 34 - 42 kDa, heparin-binding glycoprotein with potent angiogenic, mitogenic and vascular permeability-enhancing activities specific for endothelial cells. The amino acid sequence of VEGF exhibits primary structural, as well as limited amino acid sequence, homology with that of the A and B chains of PDGF. All eight cysteine residues involved in intra- and inter-chain disulfide bonds are conserved among these growth factors. A cDNA encoding a protein having a 53% amino acid sequence homology in the PDGF-like region of VEGF has been isolated from a human placental cDNA library. This protein, named placenta growth factor (PlGF), is now recognized to be a member of the VEGF family of growth factors. Based on its homology with VEGF, PlGF was also proposed to be an angiogenic factor. Two receptor tyrosine kinases have been described as putative VEGF receptors. Flt-1 (fms-like tyrosine kinase), and KDR (kinase-insert-domain-containing receptor) proteins have been shown to bind VEGF with high affinity.
In vitro, VEGF is a potent endothelial cell mitogen. In cultured endothelial cells, VEGF can activate phospholipase C and induce rapid increases of free cytosolic Ca2+. VEGF has been shown to stimulate von Willebrand factor release from endothelial cells and induce expression of tissue factor activity in endothelial cells as well as in monocytes. VEGF has also been shown to be chemotactic for monocytes and osteoblasts.In vivo, VEGF can induce angiogenesis as well as increase microvascular permeability. As a vascular permeability factor, VEGF acts directly on the endothelium and does not degranulate mast cells. It promotes extravasation of plasma fibrinogen, leading to fibrin deposition which alters the tumor extracellular matrix. The modified extracellular matrix subsequently promotes the migration of macrophages, fibroblasts and endothelial cells. Based on its in vitro and in vivo properties, VEGF is expected to play important roles in inflammation and during normal and pathological angiogenesis, a process that is associated with wound healing, embryonic development, and growth and metastasis of solid tumors. Elevated levels of VEGF have been reported in synovial fluids of rheumatoid arthritis patients and in sera from cancer patients.
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Product Summary:
The QuantiGlo Human VEGF Chemiluminescent Immunoassay is a 5.5 hour solid phase ELISA designed to measure VEGF165 levels in cell culture supernates, serum, plasma, saliva, and urine. It contains Sf 21-expressed recombinant human VEGF165 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human VEGF and recombinant human VEGF121 showed linear curves that were parallel to the standard curves obtained using the QuantiGlo kit standards. These results indicate that this kit can be used to determine relative mass values for natural human VEGF.
Precision:
Intra-Assay Precision (Precision within an assay)Four samples of known concentration were tested on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays)Four samples of known concentration were tested in separate assays to assess inter-assay precision.
The recovery of VEGF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Sample Type
Average % Recovery
Range %
Cell Culture Media (n=4)
103
95-113
Serum (n=4)
102
90-112
EDTA Plasma (n=4)
104
98-110
Heparin Plasma (n=4)
104
96-111
Urine (n=4)
99
90-106
Linearity:
To assess the linearity of the assay, samples containing or spiked with high concentrations of VEGF in various matrices were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
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