Description
Introduction: Basic fibroblast growth factor, also known as bFGF, FGF2 or FGF-beta, is a member of the fibroblast growth factor family. In normal tissue, basic fibroblast growth factor is present in basement membranes and in the subendothelial extracellular matrix of blood vessels. It stays membrane-bound as long as there is no signal peptide. It has been hypothesized that, during both wound healing of normal tissues and tumor development, the action of heparan sulfate-degrading enzymes activates bFGF, thus mediating the formation of new blood vessels, a process known as angiogenesis. Additionally, bFGF is a critical component of human embryonic stem cell culture medium; the growth factor is necessary for the cells to remain in an undifferentiated state, although the mechanisms by which it does this are poorly defined. It has been demonstrated to induce gremlin expression which in turn is known to inhibit the induction of differentiation by bone morphogenetic proteins.It is necessary in mouse-feeder cell dependent culture systems, as well as in feeder and serum-free culture systems.
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to bFGF. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP)-conjugated monoclonal antibody preparation specific for bFGF and incubated. Then substrate solution A and B are added to each well. Only those wells that contain bFGF, HRP-conjugated antibody 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 bFGF in the samples is then determined by comparing the O.D. of the samples to the standard curve