TGR5 Reporter Assay System (Human)

This Human G Protein-Coupled Bile Acid Receptor (TGR5) assay kit is an all-inclusive luciferase reporter assay system that includes, in addition to TGR5 Reporter Cells, two optimized media for use in recovering the cryopreserved cells and in diluting test samples, reference activator Lithocholic Acid (LCA), Luciferase Detection Reagent, a cell culture-ready assay plate, and a detailed protocol. INDIGO's cell-based reporter assays allow scientists to detect any biological activity that their test samples may exert against a specific receptor present in the cell. They utilize firefly luciferase reporter gene technology which provides superior precision and sensitivity. Since the receptor binding controls the expression of the luciferase reporter gene, luciferase activity in the cells can be correlated directly with the activity of the receptor. The strength of an interaction of a chemical with the target receptor is quantified using a luminometer to measure the level of light emitted. Thaw, Feed, Dose, Read Fast, reproducible, easy-to-analyze results are only four steps away Many luciferase reporter assays require the user to grow their own cells and take time to optimize the results. INDIGO's reporter cells contain the receptor of interest and the luciferase reporter gene. Reporter cells have been optimized to provide extreme sensitivity to quantify even small changes in receptor activity. With INDIGO's cell-based reporter assays, the process is as easy as Thaw, Feed, Dose, and Read. TGR5 Reporter Cells TGR5 Reporter Cells are prepared using INDIGO's proprietary CryoMite(TM) process. This proprietary cryopreservation process, enables long-term preservation of our unique reporter cells, so we can ship our cryopreserved reporter cells and assay reagents to you via overnight delivery, for your immediate use. Or, you can store the assay kits at -80°C. Once thawed, reporter cells are ready for immediate use so there is no need to take time on intermediate spin-and-wash steps, viability determinations, or cell titer adjustments. TGR5 Luciferase Reporter INDIGO's TGR5 assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set. Protein function: Receptor for bile acid. Bile acid-binding induces its internalization, activation of extracellular signal-regulated kinase and intracellular cAMP production. May be involved in the suppression of macrophage functions by bile acids. [The UniProt Consortium] TGR5 was characterized in 2002 as the first cell surface receptor for bile acids. It is encoded by the GPBAR1 gene, which is conserved among vertebrates, indicating its physiological importance. TGR5 is expressed in various organs (such as gallbladder, intestine, spleen, brown adipose tissue, and skeletal muscles) and cell types (such as macrophages, liver endothelial, and Kupffer cells). As a member of the G Protein-coupled receptor family, the topology of TGR5 displays the characteristic seven transmembrane helices, three extracellular loops contributing to ligand binding, and an intracellular carboxy tail that associates with trimeric G proteins. Upon binding to bile acid ligands, TGR5 undergoes a conformational change that triggers the activation of Gas proteins via an exchange of GDP with GTP, followed by the activation of adenylate cyclase and the production of cAMP. TGR5 functions as a metabolic regulator involved in the homeostasis of bile acids, glucose metabolism, lipid metabolism and energy expenditure. In addition, TGR5 regulates pathways related to inflammation, cancer, and liver regeneration. Interestingly, The TGR5 promoter harbors the response element for the Farnesoid X Receptor (FXR), the predominant bile acid nuclear receptor, resulting in ligand-dependent cross-signaling between FXR and TGR5. INDIGO's TGR5 Reporter Cells include the luciferase reporter gene functionally linked to tandem Cyclic AMP Response Elements (CRE) and a minimal promoter. Activated adenylate cyclase results in the production of cAMP, which binds the transcription factor CREB (cAMP Response Element-Binding Protein). Activated CREB binds to CRE sequences, seeding the formation of a complete transcription complex that drives luciferase gene expression. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in human TGR5 activity. Therefore, the principal application of this reporter assay is in the screening of test compounds to quantify any functional activity that they may exert against human TGR5.