Navigation Links
Monitoring Receptor-Mediated Changes in [Ca2+]i Using Single-Wavelength and Dual-Wavelength Indicators on the FlexStation Workstation

This probably reflects the less than optimal assay protocol and subsequent small dynamic range of the assay.


SUMMARY
In this study, ATP caused a concentration-dependent increase in [Ca2+]i in CHO cells endogenously expressing the P2Y2-receptor, with a potency similar to that reported previously. Furthermore, we have demonstrated that the FlexStation can be used to generate reliable, reproducible data for calcium mobilization assays used to monitor such Gq-coupled GPCRs.

The EC50 estimates for ATP were similar, whether the fluorescent dye Fluo-3 (0.639uM) or Fura-2 (0.639uM) or the FlexStation Calcium Assay Kit (0.651uM) was used, and were consistent between assay plates on the same day, as well as between assays analyzed on different days (less than 0.6 log unit differences). There were, however, significant differences in the dynamic ranges from the three different loading protocols, which is reflected in the corresponding Z-factors. Less than a twofold increase in signal above basal was observed when the cells were loaded with Fluo-3. The most likely cause is the subsequent wash steps in the protocol, reducing cell responsiveness. Using cells of a lower passage number and a dedicated cell-washer (Denley or similar) would probably have improved the Z-factor for Fluo-3 loaded cells. However, under these experimental conditions there was a 5-fold increase in signal observed with Fura-2, despite the cells undergoing a number of wash steps. This could be attributed to the beneficial ratiometric properties of the dye. However, the biggest fold increase in signal and Z-factor was observed with the FlexStation Calcium Assay Kit indicating that this "no-wash", homogeneous assay can lead to stronger signals, higher assay precision and, thus, improved physiological data.

In conclusion, FlexStation provides an easy to use, ver
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9

Related biology technology :

1. Versatile Reporter Vectors for Monitoring Viral Transduction
2. Monitoring transfection efficiency by green fluorescent protein (GFP) detection with the Agilent 2100 bioanalyzer
3. Monitoring Microbial Populations Using Real-Time qPCR on the MJ Research Opticon 2 System
4. Monitoring Bacterial Genetic Diversity in a Freshwater Lake Using TTGE and DNA Sequence Analysis
5. Direct Monitoring of Solid-Phase Peptide Synthesis by MALDI-TOF MS
6. Monitoring fluorescence resonance energy transfer (FRET) between GFP fusions in lysates of the yeast Saccharomyces cerevisiae using the Varian Cary Eclipse
7. New statistical algorithms for Monitoring Gene Expression on GeneChip® Probe Arrays
8. Detect Single Nucleotide Sequence Changes with Mx4000 Molecular Beacon Allelic Discrimination Kits
9. Low Abundance cDNA Cloned Using Stratagenes Human Universal cDNA Library
10. Oil-Free PCR Using the Hot Top Assembly
11. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
Post Your Comments:
(Date:8/20/2014)... succeeded in measuring vibrational motion of a single molecule ... vibration of a single molecule differs from the behaviour ... performed at the University of California, Irvine, where post-doctoral ... as a visiting fellow under professor Vartkess A. Apkarian, ... was lead by Professor Eric O. Potma. The results ...
(Date:8/20/2014)... BOSTON , Aug. 20, 2014  PAREXEL International ... it will be presenting at the Baird Healthcare Conference ... President and Chief Financial Officer, will be making a ... p.m. EDT on Wednesday, Sept. 3, 2014. ... through the "Investors" section of PAREXEL,s website at ...
(Date:8/20/2014)... improve the health of your heart, researchers have discovered. ... a standard TENS machine like those designed to relieve ... the small raised flap at the front of the ... The stimulation changed the influence of the nervous system ... can drive failing hearts too hard. , Professor Jim ...
(Date:8/19/2014)... 19, 2014 Shimadzu Scientific Instruments ... Prominence-i and Nexera-i, adding to the company’s extensive ... functionality, an intuitive operating environment, and full automation, ... efficient workflow for conventional to ultra-high-speed analysis. ... and intelligent design so users can begin building ...
Breaking Biology Technology:Seeing a molecule breathe 2PAREXEL International To Present At Baird Healthcare Conference 2'Tickling' your ear could be good for your heart 2Shimadzu’s New i-Series Integrated Liquid Chromatography Systems Provide Laboratories Wider Range of Analytical Capabilities 2Shimadzu’s New i-Series Integrated Liquid Chromatography Systems Provide Laboratories Wider Range of Analytical Capabilities 3
... Using bacteria to generate energy is a signifiant step ... University of East Anglia (UEA). Published today by ... Academy of Sciences (PNAS), the research demonstrates for ... proteins which enable bacterial cells to transfer electrical charge. ...
... 23, 2011 DATATRAK International, Inc. (OTCQX: DATA), a ... for the clinical trials industry, today announced the Company,s ... for the second consecutive year. "Being ... the hard work and dedication DATATRAK and its employees ...
... In the computer displays of medical equipment in hospitals ... major role. But a discovery reported from the University ... which have been found to change their ordering and ... low concentrations of a particular bacterial lipid, might find ...
Cached Biology Technology:Discovery opens the door to electricity from microbes 2DATATRAK Recognized as 2011 Crain's Leading EDGE Award Winner 2Liquid crystal droplets discovered to be exquisitely sensitive to an important bacterial lipid 2Liquid crystal droplets discovered to be exquisitely sensitive to an important bacterial lipid 3
(Date:8/20/2014)... Ore. -- When are athletes who have suffered concussions ... Oregon study has found that high school athletes who ... 60 days experience a significant regression in their abilities ... The regression, as seen in changes in their balance ... 19 athletes. Ten of the 12 had returned to ...
(Date:8/20/2014)... a burning spiciness that is irresistible to some, but ... are using their findings to develop a new drug ... caused by inflammation or other problems. They reported their ... clinical trials, in ACS, Journal of Medicinal Chemistry ... ago, scientists had pegged a compound called capsaicin as ...
(Date:8/20/2014)... chairs, synthetic rugs and plastic bags could one day ... rather than petroleum, scientists are now reporting. The novel ... the world deal with its agricultural and plastic waste ... , Athanassia Athanassiou, Ilker S. Bayer and colleagues at ... popularity is constantly growing. In 2012, its production reached ...
Breaking Biology News(10 mins):Recovery reversal seen in Oregon study of returning concussed athletes 2Recovery reversal seen in Oregon study of returning concussed athletes 3
... Today,s opening of a new breast health center next ... significant research opportunities for the Translational Genomics Research Institute ... will include a tumor biorepository for TGen that will ... to diagnose and treat breast cancer, which affects 1 ...
... the results are receiving rave reviews from apple lovers. ... a new apple cultivar has been introduced by scientists ... New Jersey, and Indiana experiment stations. This high-quality dessert ... S. Korban, of the Department of Natural Resources and ...
... GREECETo grow the high-quality potted plants preferred by ... affect plant growth and development. The use of ... production of compact gardenia plants; the chemicals are ... the production of lateral shoots that create aesthetically ...
Cached Biology News:Breast health center gives TGen new research opportunities 2Apple enthusiasts welcome WineCrisp 2Photoselective film proves effective for controlling height in potted gardenia plants 2
Mouse monoclonal [BC-4] to Aggrecan xxPEN ( Abpromise for all tested applications). entrezGeneID: 176 SwissProtID: P16112...
Mouse monoclonal [1.F.1] to Yersinia pestis F1 antigen ( Abpromise for all tested applications). Antigen: Purified F1 antigen of Y. pestis vaccine strain EB....
7G10 anti-Fasciclin III...
Rabbit polyclonal to Nck alpha ( Abpromise for all tested applications). entrezGeneID: 4690 SwissProtID: P16333...
Biology Products: