Human VEGF Elisa Kit

background

Vascular endothelial growth factor (VEGF), formerly known as vascular permeability factor (VPF), is a signaling protein produced by cells that stimulates blood vessel formation. Specifically, VEGF is a subfamily of growth factors, the platelet-derived growth factor family within cystine-knot growth factors. They are important signaling proteins involved in angiogenesis (de novo formation of the embryonic circulatory system) and angiogenesis (growth of blood vessels from pre-existing vasculature). All members of the VEGF family stimulate cellular responses by binding to tyrosine kinase receptors (VEGFRs) located on the cell surface, causing them to dimerize and become activated through transphosphorylation, albeit at different sites, timings and degrees. The VEGF receptor has an extracellular portion consisting of seven immunoglobulin-like domains, a transmembrane spanning region, and an intracellular portion containing a split tyrosine kinase domain.

Measuring principle

The assay employs a quantitative sandwich enzyme immunoassay technique. VEGF-specific monoclonal antibodies have been pre-coated onto microwell plates. Standards and samples are pipetted into the wells, and after incubation, the coated antibody captures any VEGF present. After extensive washing, a VEGF-specific biotin-conjugated antibody is added to detect captured VEGF protein in the sample. For signal generation, horseradish peroxidase (HRP)-conjugated streptavidin was added, followed by tetramethylbenzidine (TMB) reagent. After washing to remove any unbound combination, the enzyme conjugate is added to the wells. Color development is stopped using a solution containing sulfuric acid and the intensity of the color can be measured at 450nm, which is proportional to the amount of bound protein