PDGFRa Reporter Assay System (human)

This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Platelet-Derived Growth Factor Receptor alpha (PDGFRalpha). INDIGO's PDGFRalpha reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the Human Platelet-Derived Growth Factor Receptor alpha. In addition to PDGFRalpha Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user's test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against human PDGFRalpha. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture. Features Clear, Reproducible Results All-Inclusive Assay Systems Exceptional Cell Viability Post-Thaw Consistent Results Lot to Lot Product Specifications Target Type Growth Factor Receptor Species Human Receptor Form Native Assay Mode Antagonist Kit Components PDGFRa Reporter Cells Cell Recovery Medium (CRM) Compound Screening Medium (CSM) PDGF-CC, 20.0 ug/ml Detection Substrate Detection Buffer White, sterile, cell-culture ready assay plate Shelf Life 6 months. Target Background Platelet-derived growth factor (PDGF), the physiological activator of PDGFRs, consists of four polypeptide members: A, B, C and D. The biologically active forms of PDGF proteins are both homo-dimers and hetero-dimers of disulfide-linked polypeptides. These function to promote cell migration, proliferation, and survival. Binding of dimeric PDGF triggers conformational changes that drive the assembly of homo-dimeric (Ralpha:Ralpha, Rbeta:Rbeta) and/or hetero-dimeric (Ralpha:Rbeta) receptors, and the activation of their respective cytosolic tyrosine kinase domains. Because these reporter cells constitutively express both PDGFRalpha and PDGFRbeta, it is anticipated that all three forms of the activated receptor dimers are present The tyrosine kinase activities of activated, dimeric PDGFR's initiate intracellular signaling cascades that include RAS-MAPK, PI3-AKT, PLCgamma and STAT pathways. For example, activation of the PLCgamma pathway leads to an increase of intracellular calcium. One prominent outcome of the PDGF/PDGFR > PLCgamma pathway is that calcineurin, a calcium-dependent phosphatase, dephosphorylates and activates the transcription factor NFAT. It is PDGFRalpha signal transduction via the Ca+2calcineurin / NFAT cascade that is exploited by the reporter cells provided in this kit. INDIGO's PDGFRalpha Reporter Cells contain the luciferase reporter gene functionally linked to tandem consensus sequences of NFAT response elements upstream of a minimal promoter. Activated NFAT binds to these response elements to initiate the formation of a complete transcription complex that drives Luc gene expression. Protein function: Tyrosine-protein kinase that acts as a cell-surface receptor for PDGFA, PDGFB and PDGFC and plays an essential role in the regulation of embryonic development, cell proliferation, survival and chemotaxis. Depending on the context, promotes or inhibits cell proliferation and cell migration. Plays an important role in the differentiation of bone marrow-derived mesenchymal stem cells. Required for normal skeleton development and cephalic closure during embryonic development. Required for normal development of the mucosa lining the gastrointestinal tract, and for recruitment of mesenchymal cells and normal development of intestinal villi. Plays a role in cell migration and chemotaxis in wound healing. Plays a role in platelet activation, secretion of agonists from platelet granules, and in thrombin-induced platelet aggregation. Binding of its cognate ligands - homodimeric PDGFA, homodimeric PDGFB, heterodimers formed by PDGFA and PDGFB or homodimeric PDGFC -leads to the activation of several signaling cascades, the response depends on the nature of the bound ligand and is modulated by the formation of heterodimers between PDGFRA and PDGFRB. Phosphorylates PIK3R1, PLCG1, and PTPN11. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate, mobilization of cytosolic Ca(2+) and the activation of protein kinase C. Phosphorylates PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase, and thereby mediates activation of the AKT1 signaling pathway. Mediates activation of HRAS and of the MAP kinases MAPK1/ERK2 and/or MAPK3/ERK1. Promotes activation of STAT family members STAT1, STAT3 and STAT5A and/or STAT5B. Receptor signaling is down-regulated by protein phosphatases that dephosphorylate the receptor and its down-stream effectors, and by rapid internalization of the activated receptor. [The UniProt Consortium]