cAMP/IP-One HTplex cell-based experiment performed on Mithras LB 940 using HTRF technology

Laurence Jacquemart, Bernd Hutter

Berthold technologies, Bad Wildbad, Germany. Cisbio Bioassays, Codolet, France.

GPCRs carry information within cells via two major signaling pathways: regulation of cAMPlevels and increases in intracellular Ca2+ triggered by inositol (1,4,5) tri-phosphate (IP3). These signaling pathways are activated by the specific G protein associated with the receptor. Gs and Gi coupled receptors result in variations of cAMP while Gq coupled GPCRs activate phospholipase C (PLC) and trigger the inositol phosphate (IP) cascade. Cisbio Bioassays developed a full line of quality, ready-to-use kits capable of monitoring activation of Gs, Gi and Gq coupled receptors and recently introduced a new generation of HTRF technology incorporating Lumi4-Tb™, a new TR-FRET terbium Cryptate. Thanks to this chemistry, detection of two events at the same time in one well became possible by using 2 different acceptors a green dye (cwl = 520 nm) and a red dye (cwl = 665 nm)

HTRF® Technology, GPCR research from A to Z , HTRF microplate readers