Navigation Links
Use of FlashPlate Technology for In Vitro Measurement of [35S]-GTPγS Binding in CHO Cells Expressing the Human 5-HT1B Receptor

J. Watson and J.V. Selkirk
SmithKline Beecham Pharmaceuticals, UK

Abstract

FlashPlate is a 96-well white polystyrene microplate with scintillant-coated wells designed for highvolume, in-plate radiobinding assays. This study investigates the use of FlashPlate as a platform for the measurement of G-protein activation by means of an assay of [35S]-GTPγS binding in Chinese hamster ovary cells. Results are compared to those obtained using a conventional filtration method.

Introduction

When an external signal is received by a membrane bound receptor, a series of cellular events takes place which results in the activation of an effector molecule. Many receptors transmit this signal via G-protein activation. This response can be used as a functional measure of receptor-ligand association.

The basic mechanism behind G-protein activation involves the exchange of bound GDP for GTP. Using a radioactive, non-hydrolyzable form of GTP, such as [35S]-GTPγS, G-protein activation can be assessed by measuring the accumulation of bound radioactivity.

We investigated the use of FlashPlate technology (Basic FlashPlate, SMP200) to measure [35S]-GTPγS binding, and briefly compared it to the conventional filtration method. The study examined 5-HT activation of human 5-HT1B receptors expressed in Chinese hamster ovary (CHO) cells.

Methods
Receptor Source

A Chinese hamster ovary cell line (ACC098) expressing the human 5-HT1B receptor was used as the receptor source. Prior to [35S]-GTPγS binding assay, the CHO cells were prepared as described by Thomas, et al ., 1995, and frozen in aliquots until required.

Filtration Assay

[35
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Counting FlashPlates on the Wallac MicroBeta
2. Use of Novel FlashPlate Technology to Measure cAMP Accumulation in Chinese Hamster Ovary Cells Expressing Human -2 Adrenoreceptors
3. Use of FlashPlate Technology for In Vitro Measurement of 125I-Labeled TGF-1 Binding on Chimeric Extracellular Domain of Type II Transforming Growth Factor Receptor
4. In Vitro Measurement of the Second Messenger cAMP: RIA vs. FlashPlate
5. Evaluation of FlashPlate in a Helicase Assay
6. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
7. Enhancing Ruggedness and Full-Scan MS Sensitivity Using Ion Sweep Technology
8. Novel MicroRNA Array Technology for Sensitive miRNA Profiling
9. DNA purification from pig ears to bird blood using proven DNeasy Technology
10. Near Infrared Technology and Optical Agents For Molecular Imaging
11. In-Vitro Metabolism Studies Using Data-Dependent LC/MSn
Post Your Comments:
(Date:7/24/2008)...re/ -- The Global Probiotics Council,(GPC), a comm...ult Honsha,Co., Ltd., announced the two awardees w...rant for Probiotics Research (YIGPRO) today. The p...who will continue to contribute to,the advancement...United,States., Ajay S. Gulati, MD, an Assistant ...
(Date:7/24/2008)... Important Advance in Radiofrequency ...if., July 24 /- PEAK Surgical, Inc., ...tissue dissection system,based on a proprietary te...rug Administration has granted the company 510(k) ... use in general surgery. The PEAK Surgery System,c...
(Date:7/24/2008)...ire/ -- Sagent Pharmaceuticals, Inc.,a privately-...nced that it,has launched ceftazidime for injectio...m antibiotic. Sagent will offer ceftazidime in 1,a...ge vials. According to,IMS data, 2007 sales of inj...ximately $45.7 million., "Sagent is proud to be p...
(Date:7/24/2008)...swire-FirstCall/ -- United,Therapeutics Corporatio...lease its 2008 second quarter financial results be...., United Therapeutics will host a half-hour tele.... Eastern Time. The teleconference is accessible,b...rs dialing,1-706-679-8129. A rebroadcast of the te...
Breaking Biology Technology:Global Probiotics Council Announces First Awardees for New Young Investigator Grant for Probiotics Research 2Global Probiotics Council Announces First Awardees for New Young Investigator Grant for Probiotics Research 3Global Probiotics Council Announces First Awardees for New Young Investigator Grant for Probiotics Research 4PEAK Surgical Receives 510(k) Clearance to Market PEAK(R) Surgery System for Use in General Surgery 2PEAK Surgical Receives 510(k) Clearance to Market PEAK(R) Surgery System for Use in General Surgery 3PEAK Surgical Receives 510(k) Clearance to Market PEAK(R) Surgery System for Use in General Surgery 4Sagent Pharmaceuticals Launches Ceftazidime for Injection, USP 2
...r some people because they either dont understand ...le have their own perceptions about the value of n...e people, networking is only important when a need...People with this attitude fail to realize that net...y, their needs will be met with less effort and gr...
...book, Islands in the Clickstream is a collection... will be reprinting excerpts from selected chapter...he month. This week features an excerpt from chapt... hope you will enjoy these samples of Thieme,s wri...r with taking the moral high ground is that, once ...
...ng companies often face a dilemma as they reach a ...tional capital for growth, but are challenged by e...ditional bank financing requirements. The Madison... existing programs that fills a void for growth ca... , ,Founded in 1977 as a local non-profit developm...
...tes was hit with the largest blackout in its histo...posed holes in the grid that were previously unkno...anies, now that preparing for a similar disaster h...effects were unprecedented, shutting down a hundre...a21 in the first three minutes alone. Fifty millio...
Other Biology Technology:Dispelling the top five myths about attending networking events 2Dispelling the top five myths about attending networking events 3Dispelling the top five myths about attending networking events 4Islands in the Clickstream: Hactivism and Soul Power 2Islands in the Clickstream: Hactivism and Soul Power 3Islands in the Clickstream: Hactivism and Soul Power 4$2 million venture debt fund available for high-growth Dane County businesses 2$2 million venture debt fund available for high-growth Dane County businesses 3$2 million venture debt fund available for high-growth Dane County businesses 4One year after the 2003 blackout: Is Wisconsin prepared? 2One year after the 2003 blackout: Is Wisconsin prepared? 3
(Date:7/24/2008)...soforms from Novel Structure Proteins (NSP), a new...Sbarro Institute for Cancer Research and Molecular...e and Technology, could be involved in apoptosis o... four years ago by Nianli Sang, then a doctoral st...ssor at Thomas Jefferson University and the Cardez...
(Date:7/24/2008)...LUMBUS, Ohio -- A single typhoon in Taiwan buries ...ment -- as all the other rains in that country all... Ohio State University study published in a recent...the first ever to examine the chemistry of stream ...sea while a typhoon was happening at full force --...
(Date:7/23/2008)..., DALLAS July 24, 2008 One of the reasons peop...at they reduce their intake of fructose, a type of...ording to a researcher at UT Southwestern Medical ...ssor of clinical nutrition and lead author of a st...f Nutrition , said her team,s findings suggest tha...
(Date:7/23/2008)...UCLA "world summit" will bring together internatio...ding the U.S., Russia, China, Japan, Australia and...om July 27 to Aug. 2. , "We are bringing toge...ncient life," said summit organizer J. William Sch... Geophysics and Planetary Physics, Center for the ...
Breaking Biology News(10 mins):Novel structure proteins could play a role in apoptosis 2Study: Typhoons bury tons of carbon in the oceans 2Study: Typhoons bury tons of carbon in the oceans 3Study: Typhoons bury tons of carbon in the oceans 4Limiting fructose may boost weight loss, UT Southwestern researcher reports 2Limiting fructose may boost weight loss, UT Southwestern researcher reports 3UCLA 'world summit' to explore ancient life on Earth, and beyond 2Congressional R 26amp 3BD Caucus co chairs join scientists engineers and graduate students 13484 1Congressional R 26amp 3BD Caucus co chairs join scientists engineers and graduate students 13484 2Congressional R 26amp 3BD Caucus co chairs join scientists engineers and graduate students 13484 3United States and Mexico Renew Agreement on Cooperation in Health 13480 1United States and Mexico Renew Agreement on Cooperation in Health 13480 2Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 1Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 2Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 3Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 4Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 5Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 6Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 7Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 8Vermillion Summarizes Recent Accomplishments and 2007 Financial Results 3B Outlines 2008 Business Milestones 3713 9Watson Files Application for Generic SEASONIQUE 28R 29 13473 1Watson Files Application for Generic SEASONIQUE 28R 29 13473 2
...ble shortening of telomeres, the protective "caps"...linked to aging and physical decline. Once they ar...e to grow old. , Researchers at Salk Institute for... that the roundworm Caenorhabditis elegans succumb...are still long, and moves with a youthful spring i...
...rsity of North Carolina at Chapel Hill School of M...r Center have identified a protein that may inhibi...CIB1, or calcium and integrin-binding protein 1, w... platelet protein that may play a role in clotting...y of cellular functions that allow processes such ...
...running back Cedric Humes was able to play against...e) thanks, in part, to a prototype composite brace...ian Love, a professor of materials science and eng...omaterials class met with Mike Goforth, Virginia T...Humes, accident, which was less than three weeks b...
...tions when no medical necessity exists - Florida,...ashington, Florida increase most , The number of ... Cesarean section (C-section) rose by 36.6 percent...today by HealthGrades, the leading provider of ind...wide variation from state to state in the rate of ...
Other Biology News:Tiny roundworm's telomeres help scientists to tease apart different types of aging 2Tiny roundworm's telomeres help scientists to tease apart different types of aging 3UNC scientists discover new role for protein as fundamental inhibitor of cell movement 2Virginia Tech football player uses prototype cast 2'Patient-choice' C-section rate rises 36%: HealthGrades study 2'Patient-choice' C-section rate rises 36%: HealthGrades study 3
RABBIT ANTI HUMAN CHYMOTRYPSIN
Rat neural stem cell kits are intended for the in vitro growth and commitment to neuronal and glial lineage differentiation, of cultured fetal rat neural stem and progenitor cells expanded using FGF-
SHEEP ANTI DEHYDROEPIANDROSTERONE
Because it is manufactured at our ISO 9001–certified facilities in Eugene, Oregon, we can guarantee that FluoroPure Grade DAPI is greater than or equal to 98% pure by HPLC.
Biology Products: