Product Description and Product Data
This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Platelet-Derived Growth Factor Receptor α and β (PDGFRα/β). INDIGO’s PDGFRα/β reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the Human Platelet-Derived Growth Factor Receptor α and β. In addition to PDGFRα/β 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 PDGFRα/β. 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.
Clear, Reproducible Results
- All-Inclusive Assay Systems
- Exceptional Cell Viability Post-Thaw
- Consistent Results Lot to Lot
|Target Type||Growth Factor Receptor|
|Shelf Life||6 months|
|Shipping Requirements||Dry Ice|
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 (Rα:Rα, Rβ:Rβ) and/or hetero-dimeric (Rα:Rβ) receptors, and the activation of their respective cytosolic tyrosine kinase domains. Because these reporter cells constitutively express both PDGFRα and PDGFRβ, 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, PLCγ and STAT pathways2, 3. For example, activation of the PLCγ pathway leads to an increase of intracellular calcium4 . One prominent outcome of the PDGF/PDGFR > PLCγ pathway is that calcineurin, a calcium dependent phosphatase, dephosphorylates and activates the transcription factor NFAT4. It is PDGFR signal transduction via the Ca+2∙calcineurin / NFAT cascade that is exploited by the reporter cells in this assay
The clinical use of recombinant PDGF-BB has led to the successful treatment of chronic or diabetes-related non-healing lower extremity wounds. In addition, PDGF-BB has also been used in clinics for reducing Parkinsonian symptoms. However, dysfunctional PDGFR signaling can lead to a range of physiological disorders. For example, enhanced signaling of PDGFRs has been implicated in the pathogenesis of atherosclerosis, pulmonary fibrosis, angiogenesis, and tumorigenesis. Consequently, the PDGF receptors continue to command much interest as targets for drug development and drug safety screening.
INDIGO’s PDGFRα/β 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. PDGF activates PDGFRα/β in a dose-dependent manner, thereby triggering the Ca+2∙calcineurin/NFAT signal transduction pathway. Quantifying relative changes in luciferase activity in the treated reporter cells relative to the untreated cells provides a sensitive, dose-dependent surrogate measure of drug-induced changes in PDGFRα/β activity.
The principal application of this reporter assay is in the screening of test materials to quantify any functional interactions, either activating or inhibitory, that they may exert against PDGFRα/β, or the coupled Ca+2∙calcineurin/NFAT signal transduction pathway.
Also available as a service