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.
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.
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.
