Human Insulin-like Growth Factor-1 Receptor Reporter Assay System

This assay utilizes proprietary human cells that have been engineered to provide constitutive expression of the Human Insulin-like Growth Factor-1 Receptor referred to herein as IGF-1R. IGF-1R is a tetrameric transmembrane protein2. The extracellular α2 subunits form the ligand-binding domain, and the disulfide linked β subunits form the transmembrane domains and the intracellular tyrosine kinase domains6. Following IGF-I binding, the tyrosine kinase domains are activated and initiate intracellular signaling cascades that include RAS-MAPK pathways. For example, activation of the RAS-MAPK pathway leads to activation of ERK1/2 and subsequent phosphorylation and activation of the transcription factor Elk-17. It is IGF-1R signal transduction via the RAS-MAPKERK1/2 cascade that is exploited by the reporter cells provided in this kit.

INDIGO's IGF-1R Reporter Cells express a hybrid Elk-1transprion factor in which the native Elk-1 DNA-binding domain (DBD) has been replaced with the yeast Gal4 DBD sequence. The luciferase reporter gene is functionally linked to an upstream Gal4 Upstream Activation Sequence (UAS). When activated, Elk-1 binds to the UAS elements to initiate the formation of a complete transcription complex that drives Luc gene expression. Quantifying 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 IGF-1R activity. The principal application of this assay is in the screening of test materials to quantify any functional interactions, either activating or inhibitory, that they may exert against IGF-1R or the coupled RAS-MAPK pathway.

INDIGO’s Reporter Cells are transiently transfected and prepared as frozen stocks using a 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 intermediate treatment steps such as spin-and-rinse of cells, viability determinations or cell titer adjustments prior to assay setup.

This assay kit provides the convenience of an all-inclusive cell-based reporter assay system for IGF-1R signaling. In addition to Reporter Cells, included are an optimized culture medium for use in reviving the cryopreserved cells, a culture medium for use in preparing test sample treatments, the physiological activator Insulin, Luciferase Detection Reagents, and a cell culture-ready assay plate. [INDIGO Catalog Nos. IB29001 | IB29002]

WARNING This product is not for human or veterinary use.