Description
Principle of the assay: This assay employs a two-site sandwich ELISA to quantitative TGF-beta1 in Bovine serum, plasma. An antibody specific for TGF-beta1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any TGF-beta1 present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for TGF-beta1 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of TGF-beta1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background: TGF- beta is capable of producing a variety of effects and virtually all cell types respond to this factor in some way. The inappropriate presence of active TGF-beta1 has been implicated in a variety of pathological conditions Because of the necessity for regulating its activity tightly, TGF-beta is secreted by cells in the form of an inactive complex. This complex consists of TGF- beta 1 associated non-covalently with a protein designated the latency associated peptide (LAP). TGF- beta 1 and LAP represent components of a pro-peptide that is cleaved in a post-golgi compartment prior to secretion. LAP and TGF- beta 1 each consist of a disulfide-linked homodimer and the association of these two components renders TGF-beta1 inactive and inaccessible to anti-TGF-beta antibodies