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Human IGF-1R Reporter Assay Kit

1 x-96 well format assays
1 x-384 well format assays
1 x-96 well format assays
1 x-384 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit for the Human Insulin-like Growth Factor-1 Receptor (IGF-1R). INDIGO’s IGF-1R reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of IGF-1R. In addition to IGF-1R Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference activator, 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 activating or inhibitory, that they may exert against human IGF-1R or the coupled RAS-MAPK pathway. 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

Product Specifications

Target TypeGrowth Factor Receptor
Receptor FormNative
Assay ModeAntagonist
Kit Components
  • IGF-1R Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • IGF-1, (ref. activator; in PBS/0.1% BSA )
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Orthologs AvailableNo
Shipping RequirementsDry Ice
Storage temperature-80C


Activation dose-response assays were performed according to the protocol provided in this Technical Manual. 200 μl / well of IGF-1R Reporter Cell suspension was dispensed into the 96-well assay plate, which was then incubated for 4 hours. The concentrated stock of IGF-I (provided), IGF-II, and Insulin (Peprotech) were further diluted using CSM to produce treatment media at the desired assay concentrations. The pre-culture media were discarded from the assay wells and 200 μl per well of the prepared treatment media were dispensed (n = 3/conc.), including ‘untreated’ control wells. Following 22-hours incubation the treatment media were discarded, Luciferase Detection Reagent was added, and luminescence intensity per well was quantified. Values of average relative light units (RLU) and corresponding values of standard deviation (SD), percent coefficient of variation (%CV), Fold-Activation and Z’ were determined for each treatment concentration. GraphPad Prism software was used to perform the least-squares method of non-linear regression to plot Fold Activation vs. Log10[ng/mL] and determinate EC50 values.
IGF-1R reporter cells were co-treated with an EC80 concentration of the reference activator IGF-I (provided) and varying concentrations of the IGF-1R inhibitors Linsitinib, BMS-536924, BMS-754807 and Ceritinib (all compounds obtained from Cayman Chemical, Ann Arbor MI, USA). INDIGO’s Live Cell Multiplex (LCM) Assay confirmed that no treatment concentrations were cytotoxic (data not shown). Non-linear regression analyses of RLU vs. Log10[Inhibitor, nM] were plotted and IC50 determinations made using GraphPad Prism software.

Target Background

Insulin-like Growth Factor-1 Receptor (IGF-1R) is a Type 1 IGF receptor belonging to the receptor tyrosine kinase (RTK) family. IGF-1R activity is mediated by both IGF-1 and insulin. Upon ligand binding, the receptor is capable of activating several signal transduction pathways, including PI 3-Kinase and Ras-MAPK to regulate proliferation, migration, and inhibition of apoptosis. Although signaling pathways mediated by both IGF-I and insulin are quite similar, the biological and physiological responses are quite different between them. Activation of IGF-I/IGF-1R regulates cellular growth and proliferation, whereas insulin mediates more dramatic glycolytic responses. Several in-vivo and in-vitro studies have indicated that correlation and relationships of IGF-I/IGF-1R signaling in cancer development as well as cancer cell lines. Now clinical approaches to inhibit IGF-I/IGF-1R signaling in cancer development have been investigated using monoclonal antibodies and small molecules.

. IGF-1R is a tetrameric transmembrane protein. The extracellular α2 subunits form the ligand-binding domain, and the disulfide linked β subunits form the transmembrane domains and the intracellular tyrosine kinase domains. 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-MAPK-ERK1/2 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.

Therefore, 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.

Also available as a service

Insulin-Like Growth Factor Receptor 1

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