Human Granulocyte-Macrophage Colony-Stimulating Factor Receptor Reporter Assay System

This assay utilizes proprietary human cells that provide constitutive expression of the Human Granulocyte-Macrophage Colony-Stimulating Factor Receptor (GM-CSFR). The glycoprotein, granulocyte-macrophage colony-stimulating factor (GM-CSF) was originally defined as a growth factor that induces the differentiation and proliferation of myeloid progenitors in response to stress, infections, and cancers. The activity of GM-CSF is mediated through binding interactions with the GM-CSF Receptor (GM-CSFR; also known as CSF2R). GM-CSFR is expressed in myeloid cells and some non-hematopoietic cells as a heterodimer comprising a ligand-specific alpha subunit and a beta subunit. The beta subunit is also shared with the heteromeric IL-3 and IL-5 receptors.

INDIGO's Reporter Cells contain the luciferase reporter gene functionally linked to an engineered minimal promoter sequence with upstream tandem STAT5 genetic response element (GRE) sequences. GM-CSF activates the GM-CSFR 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 GM-CSFR.

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 GM-CSFR 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 GM-CSF as a positive control, Luciferase Detection Reagents, and a cell culture-ready assay plate. [INDIGO Catalog Nos. IB30001]

WARNING This product is not for human or veterinary use.