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Human GM-CSFR Reporter Assay Kit

SIZE SKU PRICE
1 x-96 well format assays$910 USD
SIZE SKU
1 x-96 well format assays

Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the the Human Granulocyte-Macrophage Colony-Stimulating Factor Receptor (GM-CSFR). INDIGO’s GM-CSFR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the GM-CSFR. In addition to GM-CSFR Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, 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 agonist or antagonist, that they may exert against GM-CSFR. 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.

Features

  • Clear, Reproducible Results

  • All-Inclusive Assay Systems
  • Exceptional Cell Viability Post-Thaw
  • Consistent Results Lot to Lot

Product Specifications

Target TypeGrowth Factor Receptor
SpeciesHuman
Receptor FormHybrid
Assay ModeAgonist, Antagonist
Kit Components
  • Human GM-CSFR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • GM-CSF, (ref. agonist; in DMSO)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

GM-CSFR activation dose response analyses. Activation dose-response assays were performed according to the protocol provided in the Technical Manual. 200 ul / well of GM-CSFR Reporter Cell suspension was dispensed into the 96-well assay plate, which was then incubated for 4 hours. The concentrated stock of GM-CSF (provided) was 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 ul per well of the prepared treatment media were dispensed (n = 4/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.
GM-CSFR Inhibition dose-response analyses. GM-CSFR reporter cells were co-treated with an EC80 concentration of the reference activator GM-CSF (provided) and varying concentrations of the anti-alpha subunit receptor monoclonal antibody Mavrilimumab (MedChemExpress, Cat. #HY-P99031); (n = 3 / conc.). INDIGO’s Live Cell Multiplex (LCM) Assay was performed and confirmed that no treatment concentrations were cytotoxic (data not shown). Non-linear regression analyses of RLU vs. Log10 [Mavrilimumab, ng/mL] were plotted and IC50 determination made using GraphPad Prism software

Target Background

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.

GM-CSF is known to play an important role in cancer development and progression. GM-CSF stimulates the production and maturation of neutrophils, macrophages and dendritic cells that mediate the initial host responses to cancers. However, a fine balance exists. Too little GM-CSF prevents the appropriate production of innate immune cells and subsequent activation of adaptive anti-cancer immune responses. Conversely, too much GM-CSF production can exhaust immune cells and promote cancer growth.

GM-CSF is also commonly described as a cytokine, and is known to contribute to chronic inflammatory diseases by stimulating the activation and migration of myeloid cells to inflammation sites. An imbalance in GM-CSF production and signaling has been associated with autoimmune diseases like multiple sclerosis (MS), rheumatoid arthritis (RA), myasthenia gravis (MG), inflammatory bowel disease (IBD) and systemic lupus erythematosus (SLE). Consequently, GM-CSF and its specific receptor, GM-CSFR, command considerable interest in therapeutics development and drug safety screening.

Also available as a service

Granulocyte-Macrophage Colony-Stimulating Factor Receptor (GM-CSFR)

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