Description
The TGFβ assay uses proprietary human cells that provide constitutive expression of the Human type I and type II Transforming Growth Factor beta Receptors (TGFβ I/II), both of which are transmembrane serine/threonine kinase receptors. Receptor activation results when TGFβ binds to RII, which then phosphorylates and forms a heterodimer complex with RI. The mechanisms of TGFβ-RI/RII activation and ensuing signal transduction cascade have been well studied and involve the phosphorylation and interplay of a variety of Smad regulatory proteins and transcription factors. Additionally, cross-talk between the activated TGFβ and NF-kB signal transduction pathways have been characterized. INDIGO's Reporter Cells include the luciferase reporter gene functionally linked to tandem TGFβ Response Sequences (TRS) derived from the human plasminogen activator inhibitor-1 (PAI-1) promoter, a well-characterized TGFβ responsive target gene. The TRS sequences are readily bound by activated dimeric Smad3/Smad4 to initiate formation of a transcription complex. Quantifying changes in luciferase activity in drug-treated cells versus untreated (or vehicle-treated) reporter cells provides a sensitive surrogate measure of drug-induced changes in TGFβ activity. The principal application of this assay is in the screening of compounds / drug candidates to quantify any functional activity, either agonist or antagonist, that they may exert against the human TGFβ receptor. Reporter Cells are transiently transfected and prepared as frozen stocks using INDIGO's proprietary CryoMite™ process. This cryo-preservation method allows for the immediate dispensing healthy, division-competent reporter cells into assay plates. There is no need for cumbersome intermediate treatment steps such as spin-and-rinse of cells, viability determinations, or cell titer adjustments prior to assay setup. [INDIGO Catalog Nos. IB12001-32, IB12001, IB12002]WARNING This product is not for human or veterinary use.
