Transforming Human Growth Factor Beta Receptor I/II (TGFβR)
|Product Family||Product Number||Product Description||Technical Manual|
|IB12001-32||Human TGFβR Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB12001||Human TGFβR Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB12002||Human TGFβR Reporter Assay System, 1 x 384-well format assays||Technical Manual|
Kits are offered in different assay formats to accommodate researchers' needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications. Assay systems are all inclusive, providing reporter cells, optimized growth media, media for diluting test compounds, a positive-control agonist, luciferase detection reagent, a white assay plate, a detailed protocol, and a protocol quick guide. All kits are shipped on dry ice.
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 of of 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's TGFβ assay kit provides the convenience of an all-inclusive cell-based assay system. In addition the the TGFβ Reporter Cells, provided are two optimized media for use in recovering the cryopreserved cells and for diluting test samples. Also included is the reference agonist TGF-β1, Luciferase Detection Reagents, and a cell culture-ready assay plate.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
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 activate 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. This binding leads to the activation of downstream substrates and regulatory proteins, including transcription of different target genes that function in differentiation, chemotaxis, proliferation, and activation of many immune cells. Among TGFβR's functions is the regulation of inflammatory processes, particularly in the gut. It also plays a crucial role in stem cell differentiation, as well as T-cell regulation and differentiation.
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 TGFβR 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.
Service Assays: Human
The primary application of INDIGO's cell-based nuclear receptor assays are to quantitatively assess the bioactivity of a test compound as an agonist (activator) or antagonist (inhibition of an agonist response) of a given receptor. Service assays include a positive control reference compound and 'vehicle' control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online "Request a Quote" form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.