TrkC Reporter Assay System (Human)

This Human Tropomyosin Receptor Kinase C (TrkC, NTRK3) assay kit is an all-inclusive luciferase reporter assay system that includes, in addition to TrkC Reporter Cells, two optimized media for use in recovering the cryopreserved cells and in preparing test samples, reference activator NT-3, 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. TrkC Reporter Cells TrkC 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. TrkC Luciferase Reporter INDIGO's TrkC 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 tyrosine kinase involved in nervous system and probably heart development. Upon binding of its ligand NTF3/neurotrophin-3, NTRK3 autophosphorylates and activates different signaling pathways, including the phosphatidylinositol 3-kinase/AKT and the MAPK pathways, that control cell survival and differentiation. [The UniProt Consortium] Tropomysin receptor kinases A, B and C constitute a family of receptors denoted as TrkA, TrkB, and TrkC. Trk's are single-pass transmembrane receptors that contain an extracellular ligand-binding domain, transmembrane domain, and an intracellular tyrosine kinase domain. Neurotrophins, the physiological activators of Trk receptors, comprise a group of proteins that include Nerve Growth Factor beta (NGF-beta), Neurotrophin 3 (NT-3), Neurotrophin 4 (NT-4) and Brain Derived-Neurotrophic Factor (BDNF). The active forms of these neurotrophins are disulfide-linked homodimer polypeptides. The binding of neurotrophins to Trk's triggers receptor homodimerization and autophosphorylation. The intrinsic tyrosine kinase activities of activated, dimeric Trk receptor initiate intracellular signaling cascades that include RAS-MAPK, PI3-AKT, and PLCgamma pathways. Trk's are expressed in multiple tissue types, and are primarily involved in neuronal development, neuronal proliferation, and avoidance of programmed cell death. Chromosomal rearrangements of NTRK1-3 may result in gene fusions have been clinically validated as oncogenic drivers in a wide array of human cancers. Recent studies have identified a direct link between reduced neurotrophin levels and development of cognitive illnesses like Alzheimer's disease (AD). Not surprising, the Trk receptors command much interest as targets for drug development and drug safety screening. Indeed, small molecule neurotrophin mimics that activate TrkB and TrkC have shown promise as an emerging therapeutic for AD. In addition, the FDA has recently approved Trk inhibitors for the treatment of patients with solid tumors harboring NTRK gene fusions INDIGO's TrkC Reporter Cells include the luciferase reporter gene functionally linked to tandem consensus sequences of NFAT genetic response elements upstream of a minimal promoter. Activated NFAT binds to these response elements to seed the formation of a complete transcription complex that drives Luc gene expression. Thus, quantifying changes in luciferase expression in the treated reporter cells provides a sensitive surrogate measure of the changes in human TrkC 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 TrkC, or the coupled Ca+2calcineurin / NFAT signal transduction pathway.