Intended use
This immunoassay kit allows for the in vitro quantitative
determination of human VEGF concentrations in cell culture
supernates, serum, plasma and other biological fluids.
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Introduction
Vascular endothelial growth factor (VEGF), also known as vascular
permeability factor (VPF) or vasculotropin , is a homodimeric *4
**2 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. VEGF is expressed
by numerous rodent and tumor cells, including lung adenocarcinoma,
bladder carcinoma, fibrosarcoma, HL*0 promyelocytic leukemia, GS*9L
glioma, and U**7 lymphoma cells. In normal tissues, VEGF expression
has been found in activated macrophages, keratinocytes, renal
glomerular visceral epithelium and mesangial cells, hepatocytes,
smooth muscle cells , Leydig cells ,embryonic fibroblasts and
bronchial and choroid plexus epithelium. The expression of VEGF is
upregulated by phorbol ester, TGF-a and in hypoxia. In the
conditioned media of human choriocarcinoma cells (JAR and JE*3),
the occurrence of VEGF/PlGF heterodimers has also been
observed.
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Test principle
The microtiter plate provided in this kit has been pre-coated with
an antibody specific to VEGF. Standards or samples are then added
to the appropriate microtiter plate wells with a biotin-conjugated
polyclonal antibody preparation specific for VEGF. Next, Avidin
conjugated to Horseradish Peroxidase (HRP) is added to each
microplate well and incubated. Then a TMB substrate solution is
added to each well. Only those wells that contain VEGF,
biotin-conjugated antibody and enzyme-conjugated Avidin will
exhibit a change in color. The enzyme-substrate reaction is
terminated by the addition of a sulphuric acid solution and the
color change is measured spectrophotometrically at a wavelength of
**0 nm 2 nm. The concentration of VEGF in the samples is then
determined by comparing the O.D. of the samples to the standard
curve.
