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Human G-CSFR Reporter Assay Kit

SIZE SKU PRICE
1 x-384 well format assays—
1 x-96 well format assays—
SIZE SKU
1 x-384 well format assays
1 x-96 well format assays
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Product Description and Product Data

This is an all-inclusive cell-based luciferase reporter assay kit targeting the the Human Granulocyte Colony-Stimulating Factor Receptor (G-CSFR). INDIGO’s G-CSFR reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of the G-CSFR. In addition to G-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 G-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 G-CSFR Reporter Cells
  • Cell Recovery Medium (CRM)
  • Compound Screening Medium (CSM)
  • G-CSF, (ref. agonist; in PBS/0.1%BSA)
  • Detection Substrate
  • Detection Buffer
  • White, sterile, cell-culture ready assay plate
Shelf Life6 months
Shipping RequirementsDry Ice
Storage temperature-80C

Data

G-CSFR activation dose response analyses. Activation dose-response assays were performed according to the protocol provided in the Technical Manual. 200 μl / well of G-CSFR Reporter Cell suspension was dispensed into the 96-well assay plate, which was then incubated for 4 hours. The concentrated stock of G-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 μl 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.

Target Background

Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic cytokine that regulates the viability, proliferation, and differentiation of granulocyte precursors and the function of neutrophils by signaling through the homo-dimeric granulocyte stimulating factor receptor (G-CSFR; also known as CSF3R). G-CSFR is a single transmembrane protein that has no intrinsic tyrosine kinase activity; however, upon ligand binding the receptor undergoes a conformational change leading to the activation of several downstream pathways. G-CSFR is predominantly expressed on neutrophils throughout all stages of maturation but is also present on myeloid progenitors and endothelial cells.

In chemotherapy-induced neutropenia, the bone marrow reserve of granulocytes is decreased. Exogenous G-CSF treatment can accelerate proliferation and differentiation of progenitor cells, aiding neutrophil replenishment.

G-CSF is known to play an important role in cancer development and progression. Acute myeloid leukemia (AML) and atypical chronic myelogenous leukemia (aCML) have been disorders directly related to G-CSFR mutations. In addition, G-CSF and G-CSFR are highly expressed in 90% of human gastric and colon tumors.

G-CSF is also known to contribute to chronic inflammatory diseases by stimulating the activation and migration of myeloid cells to inflammation sites. G-CSF and G-CSFR deficient mice are profoundly protected in several models of rheumatoid arthritis (RA), and antibody blockade of G-CSF has been shown to protect against disease3. Consequently, GCSF and its specific receptor, G-CSFR, command considerable interest in therapeutics development and drug safety screening.

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Granulocyte Colony-Stimulating Factor Receptor (G-CSFR)

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