Description
Introduction: Epidermal growth factor (EGF) was discovered in crude preparations of nerve growth factor prepared from mouse submaxillary glands as an activity that induced early eyelid opening, incisor eruption, hair growth inhibition, and stunting of growth when injected into newborn mice. EGF is a member of a family of growth factors that bind to the same 170 kDa receptor, including TGF-, vaccinia growth factor and amphiregulin. EGF is initially synthesized as a large (130 kDa) precursor molecule in which the mature, soluble EGF sequence (6 kDa) is located. The precursor, which functions as a source for soluble EGF, may also have a role in mediation of intercellular communication between cells displaying pro-EGF on their surfaces and cells with EGF receptors. This juxtacrine activity is also shown by a numb of other unrelated factors.Although EGF has been detected in nearly all bodily fluids, the concentration of EGF in tissues is generally low. EGF has been detected in secretory 3 cells of eccrine sweat glands. An EGF-like activity is stored in platelets and released on degranulation. In fluids such as saliva, mammary fluids and secretions, prostatic and seminal fluids, and urine, EGF concentrations are rather high.in vitroandin vivobiological effects have been attributed to EGF. In vitro, A wide variety of EGF is a mitogen for fibroblasts and endothelial cells and promotes colony formation of In vivo,EGF induces epithelial development, promotes epidermal cells in culture. angiogenesis, and inhibits gastric acid secretion. EGF has been shown to be effective in accelerating wound healing.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to EGF. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for EGF and Avidin conjugated to Horseradish Peroxidase (HRP) is added to 4 each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain EGF, 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 450 nm +/- 2 nm. The concentration of EGF in the samples is then determined by comparing the O.D. of the samples to the standard curve