Navigation Links
Cis-Reporting System Monitors Intracellular Calcium Mobilization

The NFAT cis-reporting system includes the transfection-ready pNFAT-Luc reporter plasmid and the pCIS-CK negative control plasmid.

Construction of the pNFAT-Luc Plasmid

The backbone of the 5749-base-pair pNFAT-Luc plasmid (Figure 1) is the pFR-Luc reporter plasmid of the PathDetect cis-reporting system. To this backbone, we replaced the GAL4 binding element with four direct repeats of the NFAT binding sequence (286 to 257) from the IL-2 gene promoter, the most studied and widely used NFAT binding sequence.1,2,10,11 In addition to a polyadenylation signal downstream of the luciferase gene, the pNFAT-Luc plasmid contains another polyadenylation signal in front of the NFAT binding sites and TATA box. This design minimizes transcription read through from upstream sequences and background luciferase expression. For all reporter plasmids of the PathDetect cis-reporting systems, activation of the luciferase gene indicates interaction of uncharacterized gene products, extracellular stimuli, growth factors, or drug candidates with specific enhancer elements. When a plasmid expressing the gene of interest is cotransfected into mammalian cells along with a cis-reporter plasmid, luciferase activity indicates transcription activation.

Fig.1

Testing the pNFAT-Luc Plasmid in Jurkat Cells

Under natural cellular conditions, NFAT proteins are activated by antigen stimulation of the T-cell receptor.1 Pharmacology studies have established that NFAT proteins can be activated by the protein kinase C activator phorbol ester (PMA) in combination with the calcium ionophore ionomycin, reagents that raise free intracellular calcium.2,12 When Jurk
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Cis-Reporting System Monitors Intracellular Calcium Mobilization
2. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
3. Assess the In Vivo Activation of Signal Transduction Pathways with PathDetect Reporting Systems
4. Generate Adenovirus Vectors in E. coli by Homologous Recombination with the AdEasy Adenoviral Vector System
5. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
6. Enhanced PCR Cloning System
7. prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification
8. Performance Comparisons of Commercial RT-PCR Systems
9. Signal Transduction Reporting Systems Using Cis-Acting Enhancer Elements
10. Yeast Protein Production System Features High Yields and One-Step Purification
11. Detect Released Oligosaccharides Using the Eagle Eye II Still Video Imaging System
Post Your Comments:
(Date:1/15/2014)... device," Nobel laureate Herbert Kroemer famously observed, referring ... the junctures where layers of different materials meet. ... between layers of metal oxides are becoming increasingly ... spintronics, high-temperature superconductors, ferroelectrics and multiferroics. Realizing the ...
(Date:1/15/2014)... Jan. 15, 2014   Niagara Thermal Products is ... Buffalo BioBlower Technologies LLC ("B3") as the critical thermal ... award from the Army Corps of Engineers.  B3 has ... all biological contaminants and destroys VOCs and other chemical ...
(Date:1/15/2014)... SAN JOSE, California , January 15, 2014 ... for cancer, today announced the appointment of Thomas C Reynolds ... over 20 years, development experience gained in the biotechnology industry, ... "I am delighted to welcome Tom at this transformative ...
(Date:1/15/2014)... A study has been launched to test whether ... could help to tackle the problem of obesity.  This unique ... surgery based in Stowmarket) and academics at University Campus Suffolk ... technology, which is inspired by equipment used to collect data ...
Breaking Biology Technology:A deeper look at interfaces 2Niagara Thermal Products LLC Selected as Critical Supplier for Buffalo BioBlower Air Purification Systems 2Oxford BioTherapeutics Appoints Thomas C Reynolds MD, PhD to its Board of Directors 2Oxford BioTherapeutics Appoints Thomas C Reynolds MD, PhD to its Board of Directors 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 2Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 4
... 2011  Amgen (NASDAQ: AMGN ) announced that its ... for the first quarter of 2012. This represents an approximate 29 ... The dividend will be paid on March 7, 2012 to ... on Feb. 15, 2012.  About Amgen ...
... Inc. (PDL) (NASDAQ: PDLI ) today announced that ... payment of $0.15 per share to all stockholders owning shares ... (Logo: http://photos.prnewswire.com/prnh/20110822/SF55808LOGO )   ... humanization of monoclonal antibodies and, by doing so, enabled the ...
... tune and enhance thermal conductivity a basic property generally ... engineers a new tool for managing thermal effects in smart ... devices. The finding was made by a group of ... at Vanderbilt University, and published online in the journal ...
Cached Biology Technology:Amgen Announces 2012 First Quarter Dividend 2Amgen Announces 2012 First Quarter Dividend 3New method for enhancing thermal conductivity could cool computer chips, lasers and other devices 2New method for enhancing thermal conductivity could cool computer chips, lasers and other devices 3
(Date:7/9/2014)... cell divides, it must first make a copy of its ... that occurs billions of times a day in the human ... tools that cells use to replicate DNAthe enzymes that unzip ... a clear picture of how the process works. , Now, ... to decipher what goes on at the "replication fork"the point ...
(Date:7/8/2014)... reefs and the animals that depend on them relies ... sea creature to get fatand to be flexible about ... the first study of its kind, scientists at The ... live symbiotically with algaeare better able to recover from ... keep large energy reservesmostly as fatsocked away in their ...
(Date:7/8/2014)... increased risk of severe perineal tearing during childbirth in women ... a new study published today (9 July) in BJOG: ... This study, investigates among women who have had a third ... subsequent pregnancies and the recurrence of severe perineal tears. ... in some women the tear may be more extensive. A ...
Breaking Biology News(10 mins):Rockefeller scientists first to reconstitute the DNA 'replication fork' 2Rockefeller scientists first to reconstitute the DNA 'replication fork' 3Rockefeller scientists first to reconstitute the DNA 'replication fork' 4For corals adapting to climate change, it's survival of the fattest -- and most flexible 2For corals adapting to climate change, it's survival of the fattest -- and most flexible 3For corals adapting to climate change, it's survival of the fattest -- and most flexible 4Mode of delivery following a perineal tear and recurrence rate in subsequent pregnancies 2
... of corals are likely to succeed at the expense of ... in the Cell Press journal Current Biology that ... "The good news is that, rather than experiencing wholesale ... the mix of coral species as the ocean warms and ...
... genomes, says a new study of poison frogs from ... changes over time, but the rate at which those ... Juan C. Santos of the National Evolutionary Synthesis Center ... trying to explain why some species have faster-changing genomes ...
... ARLINGTON, Va.The deafening roar of supersonic aircraft can cause ... so the Office of Naval Research (ONR) is funding ... announced April 12. "The noise problem falls into ... noise impact on the communities surrounding air bases," said ...
Cached Biology News:Under climate change, winners and losers on the coral reef 2Athletic frogs have faster-changing genomes 2ONR taps research teams to help reduce jet noise 2
... The IHC Select Immunophosphatase Secondary Detection System ... IHC Select Prediluted Primary Antibody reagents, which ... qualitative identification of antigens by light microscopy ... Secondary Detection System may also be used ...
... FLISP Assay with SR-101 Labeled Substrates ... • Accurately measure serine protease activation ... • B-Bridge's FLISP Kits utilize cell-permeable, ... (FLISPs) to measure chymotrypsin-like activity in ...
From the company that first brought you the chromatin immunoprecipitation (ChIP) kit, a new and improved kit, EZ-ChIP, is now available to make your experiments easier, quicker, and most importantly,...
The PDS-1000 He Hepta system includes the PDS-1000 He biolistic system and the Hepta adaptor....
Biology Products: