luciferase GPCR assays - (Feb/28/2011 )
I have recently started to work on GPCR cell screening assays and I am trying to implement the luciferase detection system using a CREB vector. We work with GPCR stable transfected cell lines and we intend to assay receptor activation measuring cAMP response. In the market there are several options and I have noticed that Promega has a vast array. I would like to known if anyone can give me an opinion or personal experience on the use of CREB-luc assays using mammalian stably transfected cells (or similar systems before to proceed. The available detection system that we have is a Biotek Synergy and the aim is to measure cell activation in the presence of agonist/antagonist in 96 well plates.
I hope you can find some input from others on this forum, but please do not hesitate to contact me or Promega Tecchnical Services if you are looking for any information in particular. In addition to the transcription-based luciferase reporters for CRE activation and other GPCR-related pathways, you might also be interested in the GloSensor technology which measures cAMP directly. I can point you towards citations for these system, but I expect either system can meet your needs.