Erythropoietin Receptor (EPOR)
|Product Family||Product Number||Product Description||Technical Manual|
|IB17001-32||Human EPOR Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB17001||Human EPOR Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB17002||Human EPOR Reporter Assay System, 1 x 384-well format assays|
Human Erythropoietin Receptor Assay Kit
This EPOR assay kit is an all-inclusive firefly luciferase reporter assay system that includes in addition to EPOR Reporter Cells, two optimized media for use in recovering the cryopreserved cells and for diluting test samples, the physiological activator Erythropoietin, Luciferase Detection Reagents, and a cell culture-ready assay plate.
EPOR 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 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.
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 Erythropoietin Receptor Assay Services
The primary application of INDIGO's cell-based EPOR 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 Erythropoietin Receptor Assay Background
EPOR is a member of the class I cytokine receptor superfamily. Erythropoietin (EPO), the physiological activator of EPOR, is a glycoprotein that regulates erythropoiesis. EPO, produced by the adult kidneys, activates the cell-surface EPOR to initiate signal transduction through JAK2/STAT5, and other pathways including AKT and ERK1/2. The activation of these various pathways may culminate in the activation of cytosolic targets, or in the activation of specific transcription factors and the induction of their specific target genes.
JAK2 dependent phosphorylation and activation of the transcription factor STAT5 is a prominent outcome of EPOR activation, and it is the signaling pathway exploited by the reporter cells in this assay kit. Specifically, INDIGO's Reporter Cells contain the luciferase gene functionally linked to an engineered minimal promoter sequence with upstream tandem STAT5 genetic response element (GRE) sequences. Erythropoietin activates EPOR in a dose-dependent manner, thereby triggering the JAK2/STAT signal transduction cascade. Activated STAT5 binds to its consensus GREs to initiate the formation of a complete transcription complex that drives expression of the Luc reporter gene. Therefore, quantifying changes in luciferase activity from peptide-, drug-, or antibody-treated reporter cells relative to that of 'untreated' cells provides a sensitive, dose-dependent surrogate measure of changes in the activity of EPOR.
The clinical use of recombinant human EPO has led to the successful treatment of anemia associated with conditions ranging from chronic kidney disease, chemotherapy treatment for cancer, and surgery-associated blood loss. However, side effects including hypertension, increased risk for stroke, venous thromboembolism, and death, are also risks for patients treated with hEPO and other biosimilars. This provides an opportunity for the development of additional erythropoiesis stimulating agents (ESAs) which lack these adverse effects. Additionally, though erythroid progenitor cells express the highest levels of EPOR, it is also expressed on non-erythroid cells, including neural cells, myoblasts, and adipocytes, indicative of important physiological functions of EPO outside its essential role in erythropoiesis.
The primary application of this reporter assay is to screen test materials for any functional activity, either agonistic or inhibitory, that they may exert against the human EPOR.