Tango™ Tango™ GPCR Enabling Model

Description

• The Tango™ GPCR Enabling Model provides in an easy to use system the same reagents Invitrogen uses to make the family of Tango™ GPCR cell lines.
• The Tango™ Arrestin2-UAS-bla U20S parental cell line that has the beta-lactamase (bla) reporter gene under the control of a UAS response element stably integrated as well as the beta-arrestin/TEV protease fusion protein constitutively expressed.
• The Tango™ pDest-Gal4VP16-M vector which is used for insertion of your GPCR of interest for in the Tango™ Arrestin2-UAS-bla U2OS parental cell line.
• When the Tango™ pDest-Gal4VP16-M vector containing a GPCR of interest is integrated into the parental cell line, the chimeric receptor-Gal4VP16 protein is expressed. Upon ligand binding and receptor activation the protease tagged arrestin is recruited to the receptor. The UAS response element is cleaved from the GPCR and translocates to the nucleus and drives the beta-lactamase reporter gene expression. Activation of the reporter gene provides a quantifiable measurement of the degree of interaction between the target receptor and the protease-tagged arrestin partner and is unaffected by other signaling pathways in the cell, providing an exquisitely specific readout of target receptor activation.
• Invitrogen's Tango™ GPCR Assay technology combines the benefits of the Tango™ assay platform with the highly accurate, sensitive and live-cell beta-lactamase reporter system. The Tango™ assay platform is based upon a ligand binding to a G Protein-Coupled Receptor (GPCRs) that triggers desensitization, a process mediated by the recruitment of intracellular arrestin proteins to the activated receptor. As a result, the ligand-induced activation of GPCRs may be assayed by monitoring the interaction of arrestin with the test GPCR. A major advantage of this approach is that it does not depend on the G protein signaling specificity of the target receptor.