Navigation Links
Cell lines expressing recombinant aequorin and a G-protein coupled receptor for functional screening

INTRODUCTION

Many G-protein coupled receptors (GPCRs) trigger, upon binding of an agonist, a transient increase in intracellular calcium concentration. This variation acts as an internal secondary messenger and is an important modulator of many physiological mechanisms (reviewed by Rink (1990), Tsunoda (1993) and by Santella & Carafoli (1997)). Measurement of intracellular calcium concentration in cells expressing a GPCR can thus be used to monitor the efficiency of activation of a GPCR by various compounds known or suspected to be a ligand for this GPCR.

One of the methods of choice (reviewed by Mottheakis and Ohler, 2000) for such measurements is the use of cell lines expressing a GPCR and aequorin, such as described by Sheu et al (1993) or Button et al. (1993). In this system, cells are incubated with coelenterazine, which is the co-factor of aequorin. During this incubation, coelenterazine enters the cell (it is lipophylic and readily crosses the cell membrane) and conjugates with apoaequorin to form aequorin, which is the active form of the enzyme. When the cells are then exposed to an agonist of the GPCR, intracellular calcium concentration increases. This increase leads to the activation of the catalytic activity of aequorin, which oxidizes coelenterazine and yields apoaequorin, coelenteramide, CO2 and light. The intensity of light emission is proportional to the increase in intracellular calcium in the physiological range (Rizutto et al., 1995). Thus, in this system, measurement of light emission following agonist addition reflects its ability to activate the GPCR. Because light is emitted for only 20 to 30 seconds after activation of the GPCR, recording of the emitted light must be performed during the few seconds following agonist addition to the cells. This flash-type signal reflects the transient increase in calcium concentration following GPCR stimulation.


'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. siRNA Design Guidelines
2. 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
3. Gq-coupled Receptor Assays Using FLEXstation
4. Gq-coupled receptor assays using FLEXstation: M1 muscarinic receptor
5. Gq-coupled receptor assays: a comparison of FLEXstation and FLIPR
6. The development of a CXCR2 chemokine receptorbinding assay using the LEADseeker Multimodality Imaging System
7. The development of a melanocortin MC4 receptor binding assay using the LEADseeker Multimodality Imaging System
8. The development of a melanocortin MC5 receptor binding assay using the LEADseeker Multimodality Imaging System
9. The development of a calcitonin-gene related peptide (CGRP) receptor-binding assay using the LEADseeker Multimodality Imaging System
10. Screening for potential beta 2-adrenergic receptor antagonists using CypHer5E and IN Cell Analyzer 1000
11. The use of CypHer5 for receptor internalization studies in both a range of GPCRs and a non-GPCR
Post Your Comments:
(Date:1/22/2015)... (PRWEB) January 22, 2015 Crystal Diagnostics (CDx) ... that it has received AOAC-PTM Certifications for the six non-O157 ... and O145; collectively referred to as STEC or the “Big-6”) ... colony forming unit (cfu) per 325 g of raw ground ...
(Date:1/22/2015)... zFlo Motion , a US leader in ... product from a new partner. STT-Systems has developed ... 15 years, with offerings in the optical-motion capture, 3D scanning, ... (inertial measurement unit), iSen, is opening eyes around the world. ...
(Date:1/22/2015)... 22, 2015 Controlled Substance Compliance Services ... helps companies check the legal requirements around using controlled ... China. , As their international operations expand and ... software solutions built as a result of the first ...
(Date:1/22/2015)... 2015 Diagenode, Inc., a leading ... Bioruptor® and complete solutions for epigenetics research, recently ... alleviating the need for manual processing. The new ... ChIP of histones or transcription factors and a ...
Breaking Biology Technology:Crystal Diagnostics Awarded AOAC-PTM Accreditation for the Rapid Detection of “Big 6” E.coli Food Pathogens 23D Motion Capture Just Got Easy 2Global Compliance Service for Controlled Substances to Expand to China 2Diagenode Launches Unique Semi-Automated Solution for Chromatin Immunoprecipitation 2
... N.J., Jan. 24 Aton Pharma, ... today that it has,launched an expanded, ... assistance and patient education resources, for ... ophthalmic,insert). Lacrisert is a unique preservative-free, ...
... According to a recent College,of Pharmacy study ... for minor wound care is substantially more effective ... USA300. Scientists conducting,the study compared three common over-the-counter ... community acquired MRSA. Breaks in,the skin, such as ...
... Inc. (NYSE: NTY ), (http://www.NBTY.com ), a leading global ... cast a conference call to review,fiscal first quarter results on ... 2008 at 8:30 AM (ET). A press release announcing,results for ... (ET) that,day., A live and on-demand web cast of ...
Cached Biology Technology:Aton Pharma Introduces Expanded Distribution of Lacrisert(R) (hydroxypropyl cellulose ophthalmic insert) at Hawaiian Eye 2Aton Pharma Introduces Expanded Distribution of Lacrisert(R) (hydroxypropyl cellulose ophthalmic insert) at Hawaiian Eye 3Researchers Pinpoint Best Treatment to Reduce Deadly USA300, MRSA Staph Infections 2Researchers Pinpoint Best Treatment to Reduce Deadly USA300, MRSA Staph Infections 3NBTY to Web Cast Fiscal First Quarter Conference Call 2
(Date:1/22/2015)... Infinisource has launched its new NXG series of time clocks ... a higher standard for collecting attendance and labor data and ... installation, touch screen interface and seamless connection to cloud-based iSolved, ... solution for the small to mid-size employer. The ...
(Date:12/22/2014)... Dec. 22, 2014 Research and Markets ... the "The Global Watermarking and Fingerprinting Markets" ... http://photos.prnewswire.com/prnh/20130307/600769 This insight ... watermarking and fingerprinting markets. Watermarking aims to control ...
(Date:12/19/2014)... ( http://www.researchandmarkets.com/research/86ncd6/micro_market ) has announced the addition of ... Market" report to their offering. ... market is estimated to grow at a CAGR of 3.6% ... larger share in this market, Canada ...
Breaking Biology News(10 mins):Infinisource's NXG series sets new time clock standard 2The Global Watermarking and Fingerprinting Markets 2Micro Market Monitor: North America Perimeter Security Systems Market (Intrusion Detection Sensor, Video Surveillance, Communication/Alarm and Notification, Access Control System) 2Micro Market Monitor: North America Perimeter Security Systems Market (Intrusion Detection Sensor, Video Surveillance, Communication/Alarm and Notification, Access Control System) 3
... anesthetic desflurane does not contribute to increased production of ... it can produce more of this Alzheimers associated protein. ... year, and there has been concern that anesthetic use ... Zhang, Yuanlin Dong, Rudolph Tanzi, Zhongcong Xie, and colleagues ...
... a forthcoming special issue of the Inderscience publication, ... Volume 32, Issue 4), researchers from various fields ... track the dispersal of atmospheric pollutants, particulates, and ... Stefan Institute, in Ljubljana, Slovenia, provides an editorial ...
... study mapping the evolutionary history of animals indicates that ... be inferred from fossils and studies of living animals--was ... The study, which was funded by the National Science ... 10, 2008 issue of Nature. Using new high-powered technologies ...
Cached Biology News:Biomonitoring 2And the first animal on Earth was a ... 2And the first animal on Earth was a ... 3
... depletion cocktail is tailored to ... suspensions of rat spleen cells. ... StemSep (or compatible 0.5 Tesla) ... T cell content of the ...
CleanCut agarose is used to prepare genomic DNA plugs for use in pulsed field gel electrophoresis (PFGE) applications. Supplied as a 12 ml solution of 2% agarose....
... The 96 PW is an intelligent full microplate ... all wells simultaneously. This instrument gives you the ... outstanding washing efficiency , Performance ... and excellent wash efficiency. A dispense precision of ...
... Ideal for cell culture scale-up, these ... 10 7 - 10 9 range ... bottles or spinner flasks. • High mechanical ... leak tested prior to shipping • Certified ...
Biology Products: