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DiscoverX/PathHunter® Sargramostim Bioassay Kit/1000dp(10x96-well)/93-1078Y3-00112

The PathHunter Sargramostim Bioassay is an application of the DiscoverX Dimerization Assay platform, which can be used to detect ligand-induced functional dimerization of two subunits of a receptor-dimer pair. This assay detects GM-CSF-induced heterodimerization of the CSF2RA and CSF2RB receptors. The cells have been stably engineered to co-express CSF2RB fused to PK, and CSF2RA fused to EA. Binding of GM-CSF to the cytokine-specific CSF2RA receptor leads to heterodimerization with CSF2RB, leading to activation of intracellular signaling pathways. Receptor dimerization forces the two enzyme fragments to complement, resulting in the formation of a functional b-gal enzyme. The enzyme hydrolyzes a substrate to generate a chemiluminescent signal. Sargramostim binds to CSF2RA, stimulating receptor heterodimerization with the CSF2RB receptor, leading to an increase in signal.The bioassay kit contains all reagents needed for the assay including pre-qualified cryopreserved cells, detection reagents, cell plating reagent, positive control agonist, and assay plates.
Target/Description
CSF2RB protein
Group
Cytokine receptor
Cell Background
U2OS
Species
Human
Accession Number
NM_000395.2
Agonist/Ligand
GM-CSF
Assay Measures
Dimerization
Bioassay Target Name
CSF2RB/CSF2R
Assay Qualified With
Leukine®
Drug Trademark Owner
Leukine®is a registered trademark of Genzyme Corporation (licensed)
How it works
The graph is a representative dose-response curve with Sargramostim. Using the protocol outlined in this usermanual, the data show a potent, dose-dependent stimulation of dimerization of the components of the GM-CSF receptor (CSFR2A and CSF2RB) in U2OS cells when treated with a dose response of Sargramostim.
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