LiveWare Cell Lines & Plasmid Vectors
Live cells for GPCR assays
LiveWare® is a series of cells and expression vectors that have been successfully used in drug discovery1 and screening environments to aid the study of G-protein-coupled receptors (GPCR) that do not conventionally couple through intracellular calcium. LiveWare reagents have been effectively used on the FLIPR System, in conjunction with non-Gqcoupled members of many receptors.
1Coward, P. et al. Proc. Natl. Acad. Sci. 95(1): 352-357 (1998).
Over 60% of the known G-protein-coupled receptors signal through pathways other than those which lead to an increase in intracellular calcium (i.e., other than those which couple through Gq), and as genomics reveals more G-protein-coupled receptor targets this trend continues to increase. LiveWare reagents aid the study of G-protein-coupled receptors (GPCR) that do not conventionally couple through intracellular calcium.
LiveWare is based on a series of G-protein chimeras that have been developed by Bruce Conklin and colleagues at the Gladstone Institute, UCSF. LiveWare also includes the promiscuous G-protein, Gα16. The chimeras consist of the alpha subunit of a Gq-protein complex whose 5 carboxy-terminal amino acids have been replaced with those from one of the other G-proteins (either Gαs, Gαi, Gαo, or Gαz). These amino acids are responsible for the coupling of the receptor to its G-protein. Co-expression of these chimeras with specific non-Gq-coupled receptors may result in the generation of an intracellular calcium signal upon receptor stimulation.
- LiveWare includes host cells and plasmids that contain chimeras to enable Gαs, Gαi, Gαo, or Gαz coupled GPCR receptors to signal through calcium mobilization and take advantage of the high throughput capabilities of FLIPR Calcium Assay Kits and the FLIPR Tetra System.
- LiveWare reagents have been successfully used on the FLIPR System, in conjunction with non-Gq-coupled members of chemokine, serotonin, glutamate, dopamine, opoid, vasopressin and a- and b-adrenoceptor receptor families.
- Receptor configuration options are increased as LiveWare products are available in the following configurations (plasmids are available as hygromycin and neomycin (G418)-resistant; cells are hygromycin-resistant*
- Host cells: CHO cells stably transfected with one of the G-protein chimera genes or the Gα16 gene.
- Plasmids: Replicating G-protein expression vectors.
*Each plasmid order contains one vial of hygromycin-resistance plasmid and one vial of neomycin (G418)-resistance plasmid.