Navigation Links
In Vitro Measurement of the Second Messenger cAMP: RIA vs. FlashPlate

lation Counter and the level of bound radioactivity measured.

Results

Using the RIA method, the agonist 5-HT produced a dose-dependent inhibition of forskolin-stimulated cAMP levels with a pEC50 = 7.5 0.2 and a maximum response of 84 7% inhibition (Fig. 1a). The FlashPlate technique produced a comparable 5-HT inhibition curve with a pEC50 = 7.7 0.1 and a maximum response of 84 4% (Fig. 1b). The two assays produced similar results between other parameters, such as absolute levels of cAMP measured, and intra-assay variation (Table 1). Inter-assay variation using the FlashPlates was minimal, with only 6% error on three separate determinations of the same sample (5.07 0.33 pmol cAMP/ml). However, one obvious difference between the two methods was the time required to assay the same number of samples; the FlashPlate method saved several hours compared to the RIA method.

Discussion

To date, radioimmunoassay has been an effective method to measure cAMP within samples. It is, however, a laborious technique with few options for automation and hence not conducive for use in high throughput screening (HTS) applications. The cAMP [125I] FlashPlate Assay has overcome these drawbacks: its 96-well microplate format has proven itself highly efficacious for robotic HTS, permitting cAMP measurements to be fully automated, with improvements realized in both efficiency and accuracy. The assay kit (SMP001) is comprised of fewer components than the RIA kit, and is easier to use. Additionally, the elimination of the separation step in the FlashPlate method is a major time saving advance, and again favors full automation. The results obtained from the FlashPlates are very reproducible and comparable to those obtained from RIA, and so the former can be confidently used as an alternative to the latter. The FlashPlate method is extremely sensitive
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. In-Vitro Metabolism Studies Using Data-Dependent LC/MSn
2. High Throughput siRNA Delivery In Vitro: From Cell Lines to Primary Cells
3. Your Data: Silencing Hepatitis C Virus Replication With In Vitro Transcribed siRNAs
4. Your Data: Silencing Neural Cell Adhesion Genes with In Vitro Transcribed siRNAs
5. RNArticles: RNA Amplification Used to Develop a New In Vitro HCMV Assay
6. Use of FlashPlate Technology for In Vitro Measurement of [35S]-GTPγS Binding in CHO Cells Expressing the Human 5-HT1B Receptor
7. 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
8. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
9. Cell Proliferation and Viability Measurement
10. LabVIEW for Measurement and Data Analysis
11. Measurement of proteasome inhibition in live cells in Molecular Devices microplate fluorometers
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Vitro Measurement the Second Messenger cAMP RIA FlashPlate

(Date:5/17/2013)... May 17, 2013 Topricin Pain ... Inc., has been listed among nationally recommended over-the-counter ... initiative conducted by "U.S. News" Health in collaboration ... the “No. 1 Pharmacists Recommended designation” in ... national OTC brands, and is the only homeopathic ...
(Date:5/17/2013)... Dr. Sparano is Professor of Medicine and ... Albert Einstein College of Medicine and Associate Chairman of ... He is also Associate Director for Clinical Research ... Breast Cancer Working Group, a multidisciplinary group of physicians ... also serves as Vice Chair of the National Cancer ...
(Date:5/17/2013)... May 17, 2013 Oven Industries Inc. ... The 5R6-900 temperature controller has many user-friendly benefits. Contained ... into the wall as a self-contained temperature control ... distinctive detail makes the device unique, as well as ... can also be used universally, which allows the user ...
(Date:5/17/2013)... when the transplant recipient,s immune system identifies the transplanted ... previously thought that T cells, the immune cells that ... as chemokines in order to migrate to the transplanted ... Clinical Investigation , Fadi Lakkis and colleagues at the ... stimulation of T cells is not required for migration. ...
Breaking Biology Technology:Pharmacists' Picks: Topical BioMedics' Topricin Listed on Top Recommended OTC Health Products 2Joseph A. Sparano, MD, Named Vice Chair of ECOG-ACRIN Cancer Research Group 2Joseph A. Sparano, MD, Named Vice Chair of ECOG-ACRIN Cancer Research Group 3New Temperature Control Products Announced by Oven Industries Inc. 2
... , , , Efficiency of the standard protocols (1,2) for production of , ... , Propagate M13 phage by one of the ... For producing M13 ssDNA, mix 2.5 ml plating bacteria (57 , ... more than 3 days) , with 300 ...
... , , Bacteria, gram negative , , ... href="http://www.biocompare.com/techart.asp?id=575">Actinobacillus , pleuropnomoniae , ... href="http://www.biocompare.com/techart.asp?id=576">Actinomyces , viscosus <A , ... <A , ...
... , , , , Introduction ... , , , , ... an important part of your application. Many variables must be considered, , ... evaporation, and accessory chiller—so it's no surprise to find more , ...
Cached Biology Technology:M13/Phagemid DNA Extraction Protocol 2Eppendorf Multiporator Transfection Protocols for Eukaryotic , Cells 2Eppendorf Multiporator Transfection Protocols for Eukaryotic , Cells 3Eppendorf Multiporator Transfection Protocols for Eukaryotic , Cells 4Eppendorf Multiporator Transfection Protocols for Eukaryotic , Cells 5Eppendorf Multiporator Transfection Protocols for Eukaryotic , Cells 6Eppendorf Multiporator Transfection Protocols for Eukaryotic , Cells 7Optimizing Büchi® Rotary Evaporator Applications 2Optimizing Büchi® Rotary Evaporator Applications 3Optimizing Büchi® Rotary Evaporator Applications 4Optimizing Büchi® Rotary Evaporator Applications 5Optimizing Büchi® Rotary Evaporator Applications 6Optimizing Büchi® Rotary Evaporator Applications 7Optimizing Büchi® Rotary Evaporator Applications 8
(Date:5/16/2013)... MD FASEB MARC (Maximizing Access to Research ... for The Society for Developmental Biology (SDB) 72nd ... 2013. These awards are meant to promote the ... groups into the mainstream of the basic science ... scientists at the SDB 2013 Annual Meeting. , ...
(Date:5/16/2013)... used ancient shorelines to predict the stability of today,s largest ... shoreline from three million years ago, for example when ... to be evidence of a high sea level due to ... many scientists to think that if the world,s largest ice ... the same in our modern, progressively warming world. , ...
(Date:5/16/2013)... nanoparticles developed at MIT may someday eliminate the ... constantly monitor their blood-sugar levels and inject themselves ... sense glucose levels in the body and respond ... replacing the function of pancreatic islet cells, which ... Ultimately, this type of system could ensure that ...
Breaking Biology News(10 mins):World's biggest ice sheets likely more stable than previously believed 2Nanotechnology could help fight diabetes 2Nanotechnology could help fight diabetes 3
... at the Marine Biological,Laboratory (MBL) has published ... that supports her hypothesis that, when combined, ... as BCE--can act synergistically to alter a ... have previously studied the effects,of these pollutants ...
... with Mark Musch, a longtime University of Chicago collaborator, Goldstein ... to understand how these skinny, graceful fish can swim from ... environmental change would prompt an equally drastic physiological change: Our ... other body fluids and rapidly causing death. So how do ...
... powerful,cocktail of anti-HIV drugs known as highly active antiretroviral,therapy ... finally rid the,body of the virus. But they were ... undetectable in the body ?and,impervious to attack. While HAART ... capable ?given the right opportunity ?of replicating,and wreaking havoc ...
Cached Biology News:Clam embryo study shows pollutant mixture adversely affects nerve cell development 2Clam embryo study shows pollutant mixture adversely affects nerve cell development 3Sea skate experiment sheds light on human cell transport 2Jefferson Virologists Coax HIV Out of Hiding 2
... Cell Cultures (ECACC) has been offering training ... government institutes for over seven years. We ... introduce candidates to cell culture and a ... specialist areas such as planning and management ...
... hydrophobic cover slips for in situ ... arrays on glass slides., Constant ... do not adsorb probes. , Ready-to-use-Hybri-slips ... use. , Avoid RNA degradation-Hybri-slips are ...
...
Experion spin filters are used to filter the gel matrix and gel stain solution prior to use with the Experion automated electrophoresis system. Supplied as 10 spin filters....
Biology Products: