Epidermal Growth Factor Receptor 1 (EGFR1)
|Product Family||Product Number||Product Description||Technical Manual|
|IB13001-32||Human EGFR1 Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB13001||Human EGFR1 Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB13002||Human EGFR1 Reporter Assay System, 1 x 384-well format assays||Technical Manual|
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. Assay systems are all inclusive, providing reporter cells, optimized growth media, media for diluting test compounds, a positive-control agonist, luciferase detection reagent, a white assay plate, a detailed protocol, and a protocol quick guide. All kits are shipped on dry ice.
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 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 EGFR1 assay kit provides the convenience of an all-inclusive cell-based assay system. In addition the the EGFR1 Reporter Cells, provided are two optimized media for use in recovering the cryopreserved cells and for diluting test samples. Also included is the reference agonist EGF, Luciferase Detection Reagents, and a cell culture-ready assay plate.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
EGFR1 is a single-pass transmembrane receptor, one of four members of the receptor tyrosine kinase (RTK) family. Binding interactions with extra-cellular signaling peptides such a epidermal growth factor (EGF), transforming growth factor alpha (TGFα), or amphiregulan lead to receptor dimerization and auto-phosphorylation and activation of associated intracellular signaling proteins. Interestingly, EFG Receptors demonstrate two alternative signal processing modes: one via the membrane-bound receptor and signaling at cell surface; and the other signaling through internalized receptors, nuclear translocation, and co-association with activated transcription factors to interact directly with target gene promoter sequences.
Activated EGF Receptors are known to signal through several different pathways, including those mediated by Ras, PI3K, PLC-γ, and JAK, culminating in the activation of specific transcription factors and the induction of respective target genes. phosphorylation and activation of the transcription factor STAT3 is one prominent pathway utilized by EGFR1, and it is the signaling mechanism exploited by the reporter cells in the INDIGO kit.
INDIGO's reporter cells contain the luciferase gene functionally linked to an upstream minimal promoter and tandem STAT3 genetic response element (GRE) sequences. Activated, dimeric STAT3P (or STAT3P associated with nuclear EGFR1P) bind the STAT3 GREs to initiate the formation of a complete transcription complex that drives Luc expression. 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- or antibody-induced changes in EGFR1 activity.
Considering their significant role in rapid cell proliferation, and their involvement in the progression of many types of cancers, EFGR continues to command much interest as a target for the development of novel, specific, and predominantly inhibitory drugs and antibodies. Accordingly, the primary application of this EGFR1 Reporter Assay is to screen test materials for any functional activity, either agonist or inhibitory, that they may exert aginst the EGFR1.
Service Assays: Human
The primary application of INDIGO's cell-based EGFR1 assays are to quantitatively assess the bioactivity of a test compound as an antagonist (inhibition of an agonist response) of a given 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. To initiate a Service Study, download and complete all fields of the Excel worksheet "Service Work Order" then submit the electronic file to INDIGO Customer Service.