Millennium Science cAMP Alphascreen application note

Adenosine 3′, 5′ cyclic monophosphate (cAMP) plays a critical role in the transmission of signals by functioning as a ‘second messenger’. Binding of hormones or ligands to their respective receptors can either inhibit or enhance the production of cAMP by changing the activity of the enzyme adenylate cyclase. Adenylate cyclase is a membrane-associated enzyme that catalyses the formation of cAMP from ATP. Intracellular cAMP levels, in turn, regulate enzymatic activity of numerous protein kinases, which phosphorylate specific targets setting off a cascade of cellular events. Because of the importance of cAMP as a cellular messenger, several different reading methodologies have been developed to measure cellular levels, one of which is AlphaScreen. Here we describe and demonstrate the ability of the Synergy 2 to quantitate cAMP using this methodology.

This data demonstrates the ability of the Synergy 2 Multi-Mode Microplate Reader to perform the AlphaScreen competitive cAMP assay. The Synergy 2 and Gen5 Data Analysis Software can make AlphaScreen determinations, perform any data transformation, and provide statistics and plot the data and calculate concentrations of unknown samples. The reader uses a continuous tungsten-halogen light source to excite the AlphaScreen and in conjunction with a unique two-filter pair read technique provides very good results. The competitive assay shows a large signal-change, taking place over 4-5 decades of analyte concentrations. With low sample replicate variations analysis of samples can be made with a high degree of confidence.

For the full application note, AlphaScreen Quantitation of cAMP using the Synergy 2 Multi-Mode Microplate Reader, click on link.