GeneBLAzer™ EDG3-Gα15 HEK 293T DA Assay Kit

This cell line is available as dividing cells by requesting a quote for K1234 from dividingcells@invitrogen.com

GeneBLAzer™ EDG3-Gα15 HEK 293T DA (Division Arrested) cells and EDG3-Gα15-NFAT-bla HEK 293T cells contain the human EDG3 receptor stably integrated in the GeneBLAzer™ Gα15-NFAT-bla HEK 293T cell line. GeneBLAzer™ Gα15-NFAT-bla HEK 293T cells (Cat. No. K1212) contain a beta-lactamase reporter gene under control of the Nuclear Factor of Activated T-cells (NFAT) response element, and stably express the promiscuous G-protein, Gα15. Division Arrested (DA) cells are available in two configurations: an Assay Kit (which includes cells and sufficient substrate to analyze 1 x 384-well plate), and a tube of cells sufficient to analyze 10 x 384-well plates.

DA cells are irreversibly division arrested using a low-dose treatment of Mitomycin-C, and have no apparent toxicity or change in cellular signal transduction. Both EDG3-Gα15 HEK 293T DA cells and EDG3-Gα15-NFAT-bla HEK 293T cells are functionally validated for Z'-factor and EC₅₀ concentrations of Sphingosine-1-phosphate (S1P), (Figure 1). In addition, EDG3-Gα15-NFAT-bla HEK 293T cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time (data available upon request). Additional testing data using alternate stimuli are also available.

EDG-3(Endothelial-differentiation-gene-3)/S1P-3(Sphingosine-1-Phosphate-3) is a Gq/Gi/Go/G13/G12 coupled GPCR. EDG-3 has been shown to be responsible for bradycardia that has been induced in clinical trials as a side effect of the immunosuppressive drug FTY720 that targets EDG-1. EDG-3 has also been shown to induce vasodilation in response to sphingosylphosphorylcholine (SPC), sphingosine-1-phosphate (S1P), and lysosulfatide (LSF). The EDG-3 Receptor has also been shown to play a cooperative or redundant role in angiogenesis with EDG-1