Press Release (ePRNews.com) - GöTTINGEN, Germany - Apr 13, 2016 - While fluorescent dye-based assays are powerful tools for high-throughput screening in engineered cell lines or cell lysates to show that signaling has occurred, genetically encoded sensors that measure dynamic processes in physiologically relevant cells provide a higher level of insight into how signals are regulated in both space and time. It is the regulation of signaling that controls cell function in both health and disease.
Montana Molecular’s fluorescent assays for cAMP, DAG, and PIP2 are compatible with both GPCR drug discovery and basic research. Each sensor is packaged in a BacMam vector for efficient delivery to a wide variety of cell types, including cardiomyocytes, neurons, fibroblasts, pancreatic islet cells, and osteoblasts as well as standard cell lines such as HEK293, CHO, and NIH3T3.
BacMam as a delivery vector was originally described by Boyce and Bucher in 1996. In mammalian cells, this vector, based on a baculovirus, drives strong and exclusive expression of the gene encoding the fluorescent biosensor. The amount of virus introduced into a well can be easily adjusted to control the transduction efficiency and expression levels.
cADDis cAMP Assay – Produce robust fluorescence in living cells. Signals are easily detectable on either fluorescence plate readers or imaging systems. Packaged in BacMam, a BSL-1 insect virus engineered with a mammalian promoter, for safe handling and expression in a preferred cell type.
DAG Assays – Live cell assay produces a robust change in fluorescence that can be captured on a standard fluorescence plate reader or imaging system and can be multiplexed with other fluorescent sensors emitting longer wavelength light, such as the red-fluorescent PIP2 and cAMP sensors, or the calcium sensor R-GECO.
PIP2 Sensor – PIP2 is a red, dimerization-dependent fluorescent protein sensor under the control of a CMV promoter. When PIP2 is hydrolyzed, red fluorescence decreases. This sensor can be multiplexed with the green sensors for cAMP or DAG. When co-expressed with a green dimerization-dependent monomer, GA, this sensor becomes a fluorescent protein exchange (FPX) sensor that increases in green fluorescence as the red fluorescence decreases.
Red GECO Ca2+ Sensor – The red-fluorescent, genetically encoded Ca2+ indicator for optical imaging (R-GECO) developed by Robert Campbell’s lab at University of Alberta. R-GECO can be paired with cADDis, the green-fluorescent cAMP sensor, or the green-fluorescent Upward and Downward DAG sensors to simultaneously detect both Ca2+ and cyclic AMP or Ca2+ and diacylglycerol in living cells. The baculovirus delivery system enables consistent expression in a wide variety of cell types.
ArcLight Voltage Sensor – This genetically encoded, fluorescent voltage sensing protein in a BacMam viral delivery system is a noninvasive, easy-to-use tool for studying the electrical activity of live neurons.
• Easy assay steps – No cell lysis
• BacMam delivery to most cell types
• Fluorescence imaging or plate reader detection (Z’ > 0.8)
• Physiologically relevant and cell friendly
• No forskolin or IBMX needed
• No enzymes or co-factors
• Multiplex GPCR Assays: Gs, Gq, Gi, Voltage
Oxford University researchers recently published new findings about the mechanism that mediates insulin secretion in pancreatic β cells. Montana Molecular’s DAG sensors (http://montanamolecular.com/products/products-dag-assays/) play an important role in diabetes research. In this particular project, the value of measuring multiple signaling pathways in pancreatic islets to gain a more complete understanding of diabetes drugs effects is clear.
(M. Shigeto et al.: GLP-1 stimulates insulin secretion by PKC-dependent TRPM4 and TRPM5 activation. J Clin Invest. 2015)
Curious about new fluorescent biosensors to measure Gq, Gs, and Gi signaling in living cells? Need to make simultaneous measurements of multiple second messengers? Visit Montana Molecular at Experimental Biology (http://experimentalbiology.org/2016/Home.aspx) 2016 to learn more about our new generation of red and green fluorescent assays for live cell discovery.
San Diego Convention Center
April 2-6, 2016
MoBiTec GmbH, Germany, is a certified agent for Montana Molecular in Europe. For details see http://www.mobitec.com
About Montana Molecular
Montana Molecular develops genetically encoded fluorescent biosensors and probes for cell-based assays and live cell imaging. Montana Molecular products are used for drug discovery and to observe and measure cell signaling processes.
Montana Molecular’s scientific team is focused on creating breakthrough innovations that empower both basic research scientists and drug discovery teams. The company is committed to building a sustainable biotech industry in the state of Montana, providing good jobs in a safe and healthy environment.
About MoBiTec GmbH
MoBiTec GmbH (Goettingen, Germany) is a privately held company (founded in 1987) that offers research tools for molecular and cell biology. Products include DNA vectors for cloning and expression, cell transfection reagents and cell culture tools, immobilized and soluble enzymes, products for genomics and proteomics research, numerous antibodies and recombinant proteins, superior fluorescence reagents and kits, affinity chromatography products, as well as general laboratory equipment.
In parallel to its own product lines, MoBiTec distributes products from international companies in Germany. MoBiTec products are distributed worldwide, in Germany from their home office, in other countries by distributors.
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