Navigation Links
Development of a sensitive enzyme fragment complementation assay for cyclic adenosine 3', 5' monophosphate and its validation for pharmacological screening at G protein-coupled receptors

Key words: cAMP • GPCR enzyme fragment complementation LEADseeker assay miniaturization


G protein-coupled receptors (GPCRs) are the largest family of transmembrane receptors and are responsible for the transduction of a diverse range of extracellular signals. Up to 40% of clinically marketed drugs are active at this receptor family; however, these drugs exhibit their activity at less than 10% of known GPCRs. Thus, sensitive high-throughput GPCR screening assays are required to associate the many orphan GPCRs with disease to potentially identify novel pharmaceutical agents.

Here, we describe an assay for screening at GPCRs based on enzyme fragment complementation (EFC) technology and cyclic adenosine 3’, 5’ monophosphate (cAMP) measurement. The data show the cAMP II HitHunter Chemiluminescence Assay has a sensitivity of ~ 1 nM when carried out in 1536-well microplates and imaged using the LEADseeker™ Multimodality Imaging System.


Materials
Products used

LEADseeker Multimodality Imaging System 18-1140-71

cAMP II HitHunter Chemiluminescence 90-0032-03
Assay, 1536 wells


Other materials required
Krebs-Henseleit buffer

Phosphate buffered saline (PBS) solution

Dulbecco’s Modified Eagle’s Medium

DMSO

Ethanol

Greiner Bio-one Lumitrac™ 200 microplates, 1536-wells

Biomek™ FX Laboratory Automated Workstation (Beckman Coulter)

CyBi™-well (Cy-Bio)

Chinese hamster ovary (CHO-K1)
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. The let-7 microRNA Family Regulates RAS: Implications for Development and Oncogenesis
2. Rapid Method Development With the BioLogic DuoFlow Chromatography System for the Purification of His-Tagged Proteins
3. A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis
4. Development of a Multiplex Bead-Based Assay for Antibody Screening of a Nonhuman Primate Colony on the Bio-Plex System
5. Development of a histamine H1 receptor binding assay using the LEADseeker Multimodality Imaging System
6. Development of an NK2 neuropeptide receptor binding assay using LEADseeker Multimodality Imaging System
7. Development of an adrenergic α2a receptor binding assay using the LEADseeker Multimodality Imaging System
8. Development of a CRF1 neuropeptide receptor binding assay using LEADseeker Multimodality Imaging System
9. Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets.
10. Screening for β2-adrenergic receptor agonists using the pH-sensitive dye,CypHer5, and the IN Cell Analyzer 3000
11. The combination of monolithic columns and modern HPLC/MS/MS instrumentation for high chromatographic throughput and sensitive detection of beta-agonists
Post Your Comments:
(Date:10/20/2014)... (PRWEB) October 20, 2014 The ... (Medication Dispensing Systems, Packaging and Labeling Systems, Table-top ... Automation) - Global Forecasts & Trends to 2019” ... and opportunities in North America, Europe, Asia-Pacific, and ... 101 market tables and 30 figures spread through ...
(Date:10/20/2014)... 20, 2014 Asterias Biotherapeutics, Inc. (NYSE ... signed a Notice of Grant Award (NGA) with ... October 1, 2014.  The NGA provides for the ... release of additional grant funds pursuant to the ... clinical development of Asterias, product, AST-OPC1. The grant ...
(Date:10/20/2014)... , Oct. 20, 2014  GenVec, Inc. (NASDAQ: ... Horovitz , Ph.D., from its board of directors effective on ... August 2003, and served as its chairman from June 2006 ... the Nominating and Corporate Governance and Audit Committees of the ... than a decade of dedicated service to GenVec, and its ...
(Date:10/20/2014)... 20, 2014 Earle Martin ... announced today that Ellen Teplitzky, an experienced attorney ... the pharmaceutical industry, has joined the firm as ... legal services practice. NDA Partners provides legal ... and testimony, to top law firms and their ...
Breaking Biology Technology:Pharmacy Automation Systems Market worth $4,566.2 Million by 2019 - New Research Report by MarketsandMarkets 2Pharmacy Automation Systems Market worth $4,566.2 Million by 2019 - New Research Report by MarketsandMarkets 3Pharmacy Automation Systems Market worth $4,566.2 Million by 2019 - New Research Report by MarketsandMarkets 4Asterias Biotherapeutics Announces Notice of Grant Award with CIRM for Phase 1/2a Clinical Trial of AST-OPC1 in Complete Cervical Spinal Cord Injury 2Asterias Biotherapeutics Announces Notice of Grant Award with CIRM for Phase 1/2a Clinical Trial of AST-OPC1 in Complete Cervical Spinal Cord Injury 3Asterias Biotherapeutics Announces Notice of Grant Award with CIRM for Phase 1/2a Clinical Trial of AST-OPC1 in Complete Cervical Spinal Cord Injury 4Zola P. Horovitz To Retire From GenVec Board 2NDA Partners Appoints Ellen Teplitzky, JD as Director of its Legal Services Practice 2
... to the biotech community to have a major research facility ... the Wisconsin Life Sciences and Venture Conference program ... conference will be held November 16 and 17. , ,The ... companies, 11 from Wisconsin, will strut their stuff in front ...
... to those risk-addicted Californians: Saddled with a debt running into ... their credit cards and pinch pennies. They borrow even more ... happened last week when California voters overwhelmingly approved Proposition 71, ... in bonds to pay for human embryonic stem cell research. ...
... Wisconsin-Madison Board of Regents has approved construction of a new ... and Clinics and the Waisman Center . The ... from the State Building Commission on November 17. , ,The ... an imaging center and animal quarters. The facility is designed ...
Cached Biology Technology:Life-sciences conference to feature novel research with clinical potential 2California stem cell initiative should wake up Washington and Madison 2California stem cell initiative should wake up Washington and Madison 3
(Date:10/17/2014)... regarding Ebola is critical and pertinent for practicing physicians ... outbreak and epidemic. The Journal, Disaster Medicine and ... Ebola Virus and Public Health, to surround the public, ... critical societal moment. , On October 17, the ... . The primer was prepared by Dr. Eric Toner, ...
(Date:10/17/2014)... University of Copenhagen have shown for the first time ... Cystic fibrosis patients, giving them the opportunity to get ... infections. , The study also discovered the bacterial growth ... was halted or slowed down by the immune cells. ... oxygen and helped "suffocate" the bacteria, forcing the bacteria ...
(Date:10/16/2014)... constantly wages war on the human body. Battles ... stalemate. In pancreatic cancer, this stalemate—known as tumor ... aggressively malignant, a phenomena that is poorly understood. ... the laboratory of Salvatore Torquato, a Professor of ... conditions surrounding tumor dormancy and the switch to ...
Breaking Biology News(10 mins):Scientists opens black box on bacterial growth in cystic fibrosis lung infection 2Modeling tumor dormancy 2
... seen amongst individual species within the animal kingdom evolved ... effectively serve as medical research models because humans and ... key to morphological and behavioral complexity, a growing body ... expression by a family of DNA-binding proteins called "transcription ...
... AUSTIN, Texas Researchers at The University of Texas at ... in the folic acid metabolic pathway leads to neural tube ... known for several decades that folic acid supplementation dramatically reduces ... and anencephaly, which are among the most common birth defects. ...
... press release is available in German . ... at the Max Planck Institute for Developmental Biology in ... to identify neuronal correlates of behaviour by comparing maps ... different behaviour. They compared the pharyngeal nervous systems of ...
Cached Biology News:New key to organism complexity identified 2New key to organism complexity identified 3New key to organism complexity identified 4Lack of key enzyme in the metabolism of folic acid leads to birth defects 2The neurobiological consequence of predating or grazing 2The neurobiological consequence of predating or grazing 3
... Pichia Protocols is the first all-inclusive collection ... Pichia pastoris Expression System. Methods developed by ... as detailed, step-by-step protocols. This book will ... vector, through strain construction and analysis, to ...
... Immunogen: Synthetic peptide derived from the C-terminal ... growth factor D iris-expressed growth factor spinal ... mouse and rat by one amino acid ... kDa human mouse and rat PDGF-D proteins. ...
...
...
Biology Products: