Science Catalog 2012 Life Cell Signaling GPCR Assays cAMP-Glo™ Assay Product cAMP-Glo™ Assay Size Cat.# Price (Fr) 300 assays V1501 486.00 3,000 assays V1502 2916.00 30,000 assays V1503 Pls. Enq. For Research Use Only. Not for Use in Diagnostic Procedures. Description: The cAMP-Glo™ Assay is a homogeneous, bioluminescent and high-throughput assay for measuring cAMP levels in cells. The cAMP-Glo™ Assay monitors cAMP production in cells in response to the effects of test compounds on G protein-coupled receptors (GPCR). GPCRs that couple with adenylate cyclase will increase or decrease intracellular cAMP. The assay is based on the principle that cyclic AMP (cAMP) stimulates protein kinase A (PKA) holoenzyme activity, decreasing available ATP and leading to decreased light production in a coupled luciferase reaction. The cAMP-Glo™ Assay can be performed in 96-, 384- or 1536-well plates. The cells are induced with a test compound for an appropriate period of time to modulate cAMP levels. After induction, cells are lysed to release cAMP, then the cAMP detection solution, which contains protein kinase A, is added. The Kinase-Glo® Reagent is then added to terminate the protein kinase A reaction and detect the remaining ATP via a luciferase reaction. Plates are read using a microplate-reading luminometer. Luminescence can be correlated to the cAMP concentrations by using a cAMP standard curve. The half-life for the luminescent signal is greater than 4 hours. This extended signal half-life eliminates the need for luminometers with injectors and allows batch-mode processing of multiple plates. Features: Fast and Easy to Use: • Assay can be completed in approximately 45 minutes. • Homogeneous. • Two steps following lysis of cells. Excellent Signal-to-Noise Ratios: • Best signal:background ratio of all the cAMP assays. • Signal:Background >200 (with cAMP), >15 (on cells). • Easily scalable to 1536-well plate formats and beyond. Proven Luminescent Technology: • Powered by Ultra-Glo™ Recombinant Luciferase. • No interference by fluorescent compounds. • Nonradioactive. Storage Conditions: Store the system at –20°C. Once prepared, the cAMP detection solution (cAMP-Glo™ Reaction Buffer with Protein Kinase A) should not be frozen. Once prepared, the Kinase-Glo® Reagent should be dispensed into aliquots and stored at –20°C. See the product label for the expiration date. Protocol cAMP-Glo™ Assay Technical Bulletin Part# TB357 Phosphorylated protein kinase A substrate ADP P cAMP RR C C ATP luciferin + O2 luciferase Active protein kinase A Protein kinase A substrate oxyluciferin + AMP + Light Schematic diagram of cAMP production in cells and the cAMP-Glo™ Assay. cAMP RR CC Inactive protein kinase A holoenzyme β β γ γ G protein cAMP ATP α Ligand G protein-coupled receptor Inactive adenylate cyclase GDP Active adenylate cyclase α GTP 48 For complete and up-to-date product information visit: www.promega.com/catalog 6127 MA Pagina 51
Pagina 53Voor catalogi, online nieuwsbrieven en magazines zie het Online Touch beheersysteem systeem. Met de mogelijkheid voor een webshop in uw clubmagazines.
Promega Switzerland - Life Sciences Catalog 2012 Lees publicatie 100Home