Human VEGF-R1 ELISA Kit
The sVEGF-R1 ELISA is an enzyme-linked immunosorbent assay for quantitative detection of soluble human vascular endothelial Growth Factor Receptor-1 in cell culture supernatants, human serum, plasma or other body fluids.
Soluble VEGF-R1 (sFLT-1) is a naturally occurring endogenous form of the VEGF-R1 and was originally found in the supernatant of human vascular endothelial cells. It is generated by differential splicing of the flt-1 gene. In vitro sVEGF-R1 is used to inhibit VEGF-A mediated signals in endothelial cells and in vivo it can be used to block physiological angiogenesis in several organs, e.g. in the ovary or in bones. Tumor cells transfected with the flt-1 gene are growth restricted in vivo because of the limitation in developing tumor blood vessels via VEGF-A signalling. Very recent studies have shown that this molecule is present endogenously at ng/ml concentrations in biologicals fluids of normal human subjects or in the conditioned media of FLT-1 positive cell types. The measurement of sFLT-1 in a variety of clinical conditions may open up new insights in health and disease.
An anti-sVEGF-R1 coating antibody is adsorbed onto microwells.
sVEGF-R1 present in the sample or standard binds to antibodies adsorbed to the microwells; a biotin conjugated anti-sVEGF-R1 antibody is added and binds to sVEGF-R1 captured by the first antibody.
Following incubation unbound biotin conjugated antisVEGF-R1 is removed during a wash step. StreptavidinHRP is added and binds to the biotin conjugated antisVEGF-R1. Following incubation unbound Streptavidin-HRP is removed during a wash step, and substrate solution reactive with HRP is added to the wells.
A coloured product is formed in proportion to the amount of sVEGF-R1 present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450 nm. A standard curve is prepared from seven sVEGF-R1 standard dilutions and sVEGF-R1 sample concentration determined.