Platelet-Derived Growth Factor Receptor α and β (PDGFRα/β)
|Product Family||Product Number||Product Description||Technical Manual|
|IB23001-32||Human PDGFRα/β Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB23001||Human PDGFRα/β Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB23002||Human PDGFRα/β Reporter Assay System, 1 x 384-well format assays|
Platelet-Derived Growth Factor Receptors α and β Reporter Assay Kit
This Human Platelet-Derived Growth Factor Receptors α and β (PDGFRα/β) reporter assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to PDGFRα/β Reporter Cells, an optimized culture medium for use in reviving the cryopreserved cells, a culture medium for use in preparing test sample treatments, the reference physiological activator PDGF-BB, Luciferase Detection Reagents, and a cell culture-ready assay plate.
PDGFRα/β 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 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 PDGFRα/β reporter 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.
PDGFRα/β reporter assay 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.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
Human Platelet-Derived Growth Factor Receptors α and β Reporter Assay Services
The primary application of INDIGO's cell-based PDGFRα/β assays are to quantitatively assess the bioactivity of a test compound as an agonist or antagonist of the 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.
Human Platelet-Derived Growth Factor Receptors α and β Reporter Assay 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 (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.
Synonyms: PDGFR, PDGFR alpha, PDGFR beta, Human Platelet-Derived Growth Factor Receptor alpha, Human Platelet-Derived Growth Factor Receptor beta
Human Platelet-Derived Growth Factor Receptors α and β Reporter Assay Research Areas
Wound Healing, Parkinson's Disease, Atherosclerosis, Cancer