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Promega Corporation

GloSensor™ Technology

Intracellular Detection of Signal Transduction in Living Cells

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GloSensor™ represents a platform technology of biosensors for the intracellular detection of signal transduction in living cells. Promega scientists developed the intracellular biosensor concept using directed evolution resulting in a suite of novel constructs that are easier to use and more sensitive than any other product on the market.

pdf_icon Want to Know More? Download the Technical Update
GloSensor™ Technology Overview, November 2011

The overview presents the GloSensor concept, recent performance data, and comparison against time-resolved fluorescence and an immunoassay.

Product Information

Design Concept for a cAMP Biosensor

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The conformational change of an analyte binding domain modulates luminescence, e.g., cAMP binding to RIIβB promotes increased luminescence.

The GloSensor™ cAMP Assay is a 'zero step', nonlytic, live-cell assay ideal for kinetic and modulation studies of signaling through cAMP. The simple protocol is easily scalable to your throughput needs.

Simple, Scalable Protocol

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  • Pre-equilibrate cells with GloSensor™ cAMP Reagent for ~2 hours.
  • Treat with specific agonists/antagonists or library compounds.
  • Measure luminescence after 15-30 minutes.

View complete protocol

Accessing the Technology

The GloSensor™ cAMP Assay comprises three components of which two are required to perform the assay; the Reagent in combination with the Cell Line or the Plasmid. GloSensor™ cAMP Reagent is required per a specific Limited Use Label License. For more information on licensing GloSensor Technology, please contact us.

References

Allen, JA et al. Discovery of β-Arrestin-Biased Dopamine D2 Ligands for Probing Signal Transduction Pathways Essential for Antipsychotic Efficacy. PNAS 108 (45): 18488-18493, 2011 (DOI: 10.1073/pnas.1104807108)

Pantel, J et al. Development of a High Throughput Screen for Allosteric Modulators of 3 Melanocortin-4 Receptor Signaling Using a Real Time cAMP Assay. Euro J Pharmacol 660: 139-147, 2011 (10.1016/j.ejphar.2011.01.031)

Binkowski, B et al. A Luminescent Biosensor with Increased Dynamic Range for Intracellular cAMP. ACS Chem Biol 6: 1193–1197, 2011 (dx.doi.org/10.1021/cb200248h)

Buccioni, M, Marucci, G et al. Innovative functional cAMP assay for studying G protein-coupled receptors: application to the pharmacological characterization of GPR17. Purinergic Signalling Publ online 20 July 2011 (DOI 10.1007/s11302-011-9245-8)

Rajagopal, S et al. Quantifying Ligand Bias at Seven-Transmembrane Receptors. Mol Pharmacol 80(3):367-77, 2011 (DOI:10.1124/mol.111.072801)

Oka, T et al. Bioluminescence Technologies to Detect Calicivirus Protease Activity in Cell-free System and in Infected Cells. Antiviral Research 90: 9–16, 2011 (DOI:10.1016/j.antiviral.2011.02.002)

Nobles, KN et al. Distinct Phosphorylation Sites on the β2-Adrenergic Receptor Establish a Barcode That Encodes Differential Functions of β-Arrestin Science Signaling 4 (185), ra51, 2011 (DOI: 10.1126/scisignal.2001707)

Binkowski, B, Fan, F, and Wood, KV Luminescent Biosensors for Real-Time Monitoring of Intracellular cAMP, In: Signal Transduction Protocols, Series: Methods in Molecular Biology, Vol. 756, 263-271, Luttrell, LM and Ferguson, SSG, Eds, 2011 (ISBN 978-1-61779-159-8)

Hill, S, Williams, C, and May, Lauren Insights into GPCR pharmacology from the measurement of changes in intracellular cyclic AMP; advantages and pitfalls of differing methodologies. Brit J Pharmacol 161(6): 1266-1275, 2010 (DOI: 10.1111/j.1476-5381.2010.00779.x)

Kimple, AJ et al. Structural Determinants of G-protein a Subunit Selectivity by Regulator of G-protein Signaling 2 (RGS2). J Biol Chem 284: 19402-19411, 2009 (DOI: 10.1074/jbc.M109.024711)

Binkowski, B et al. Engineered Luciferases for Molecular Sensing in Living Cells. Curr Opin Biotechnol 20(1): 14-18, 2009 (DOI:10.1016/j.copbio.2009.02.013)

Fan, F, Binkowski, B et al. Novel Genetically Encoded Biosensors Using Firefly Luciferase. ACS Chem Biol 3 (6): 346–351, 2008

Wigdal, S et al. A Novel Bioluminescent Protease Assay Using Engineered Firefly Luciferase. Curr Chem Genomics 1: 94-106, 2008

GloSensor is a trademark of Promega Corporation.

Limited Use Label License

BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE STATEMENT. If the researcher is not willing to accept the conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide the researcher with a full refund. NOT FOR USE IN LIVE ANIMALS— For use in live animals contact Promega regarding the availability of licenses. Researchers may use this product and its derivatives in their own research solely for the luminescent determination of cAMP levels in vitro or within cells (excluding uses in living animals). Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence encoding the GloSensor™ cAMP Reporter of this product (i.e., the bioluminescent chimeric protein comprising luciferase and cAMP-binding domains). Researchers may transfer derivatives to others provided that recipients agree to be bound by the conditions of this limited use statement prior to such transfer. No other use or transfer of this product or derivatives is authorized without the prior express written consent of Promega. In addition, researchers must either: (1) use the GloSensor™ cAMP Reagent purchased from Promega for all determinations of luminescence activity of this product and derivatives; or (2) contact Promega to obtain a license for use of the product and derivatives in conjunction with luminescent assay reagents not purchased from Promega. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE WITH REGARDS TO THE PRODUCT. The terms of this agreement shall be governed under the laws of the State of Wisconsin, USA.

For additional licensing questions, please contact Promega.

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Scientists at Your Service

Scientists at Your Service

We offer a range of services to help you succeed using Promega technologies. From product training to set up of automated systems and development of custom applications—our scientific support goes beyond the basics.

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