Researchers Describe a Novel In-Cell Western Assay for GPCR Mediated cAMP Response Element Binding Protein Phosphorylation

Lincoln, NE - June, 09 2006 - Researchers at Neurocrine Biosciences Inc., San Diego, CA have described a novel In-Cell Western (ICW) assay for G-Protein-Coupled Receptor–Mediated cAMP response element binding protein phosphorylation.Currently one of the methods used to determine the downstream phosphorylation is done using radio ligand binding affinities assay. In this paper, the author’s describe a high throughput ICW assay approach for the measurement of CREB phosphorylation. This, in turn can, be used to assess GPCR activity in whole cells and is more amenable to the throughput required for the purposes of drug discovery.

Using the ICW, the authors demonstrate the functional potency of adenosine2A (A2A) receptor antagonists and the functional antagonism achieved using the more upstream readout of cAMP accumulation in PC-12 cells. The assay is performed in a 96 micro-well plate to accommodate eight 12- point dose response curves.

This assay has a distinct advantage over running Western Blots, as it would take over 10 blots to achieve the same throughput and the researcher would have to deal with loading and transfer errors.

To read the abstract of this publication, please click here.

The Odyssey Infrared Imaging System is uniquely equipped with two infrared channels for direct fluorescence detection on membranes with sensitivity that is equal to or better than chemiluminescence. The superior signal-to-noise ratio produced using near infrared dyes produces results not possible with traditional fluorescence-based imaging systems. LI-COR infrared systems are noted for providing superior imaging for a growing list of applications including Western blots, In-Cell Western assays, EMSA's, gel documentation and in vivo imaging.

For more information contact:

D. Doc Chaves
Marketing Communications Manager
LI-COR Biosciences
4647 Superior Street
Lincoln, Nebraska 68504
402.467.0750
doc.chaves@licor.com