The Human TGFβ Antibody Pair Kit comes with pre-matched antibody pairs, standards, and streptavidin-HRP to develop your own enzyme linked immunosorbent assays (ELISA) to detect and quantify protein levels of human TGFβ. Buffer reagents needed to complete the ELISA reaction are sold separate as a Buffer Kit for Antibody Pairs (Catalog No. CNB0011). The Assay Buffer included in this Buffer Kit can be used as a blocking reagent for ELISA plates as well as a diluent for ELISA standards and samples, detection antibody, and HRP conjugate.
Principle of the method
ELISAs are designed to measure the amount of the target bound between a matched antibody pair. A target-specific antibody is coated to the bottom of the wells of a microplate, which is an overnight process. Samples, standards, or controls are then added into these wells and bind to the immobilized (capture) antibody. A sandwich is formed by the addition of the second (detector) antibody, a substrate solution is added that reacts with the enzyme-antibody-target complex to produce measurable signal. The intensity of this signal is directly proportional to the concentration of target present in the original specimen.
TGF beta-1 (TGFB1, Transforming Growth Factor Beta 1) is a polypeptide member of the transforming growth factor beta superfamily of cytokines, found almost ubiquitously in tissues. Transforming growth factor (TGF)-b is stored in the extracellular matrix as a latent complex with its pro-domain. Activation of TGF beta-1 requires the binding of aV integrin to an RGD sequence in the prodomain and exertion of force on this domain, which is held in the extracellular matrix by latent TGF-b binding proteins. Latent forms are complexes of TGF beta-1, an amino-terminal portion of the TGF-beta precursor, designated TGF-LAP (TGF-latency associated peptide), and a specific binding protein, known as LTBP. TGF beta-1 helps regulates proliferation, differentiation, adhesion, migration in many cell types. Many cells have TGF beta receptors, and the protein positively and negatively regulates many other growth factors. TGF beta-1 is cleaved into a latency-associated peptide and a mature TGF beta-1 peptide, and is found in either a latent form composed of a TGFB1 homodimer, a LAP homodimer, and a latent TGFB1-binding protein, or in an active form composed of a TGF beta-1 homodimer. The mature peptide may also form heterodimers with other TGF beta family members. The gene for TGF beta-1 is frequently upregulated in tumor cells, and mutations in this gene result in Camurati-Engelmann disease and cystic fibrosis.
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