Navigation Links
Stable HeLa Luciferase Reporter Cell Lines Expressing GAL4 Fusion,,,Transactivators

tems mentioned above and had the lowest background of luciferase activity.

Next, in separate experiments, we transfected the HLR cell line with the fusion trans-activator pFA2-CREB, pFA2-Elk1, or pFA2-cJun plasmids. After performing successive rounds of selection with G418, we chose the clones that gave the best response when transfected with the positive control vectors from the PathDetect kits. These clones were then subjected to more extensive analysis.

Testing Stable Cell Lines

Fig.2

Genomic DNA from each stable clone was prepared and used as a template for PCR analysis to confirm that each fusion trans-activator plasmid was stably incorporated into the genomes of the selected clones. To amplify the activation domain for each reporter cell line, we used one primer specific to the GAL4 DNA-binding domain and another primer specific to each of the activation domains. Each primer set successfully amplified a product of the expected size. Analysis of HLR-CREB and HLR-Elk1 cell lines are shown in Figure 2. To further confirm the activity and specificity of these cell lines, we transfected each cell line with plasmids encoding known activators of each pathway. Upon luciferase assay of cell lysates, each cell line showed significant luciferase expression above background (Figure 3).

Fig.3

Additional testing included an analysis of the activation of luciferase activity in these stable cell lines by extracellular stimuli. For example, HLR-Elk1 cells were treated with known MAPK activators, including epidermal growth factor (EGF) and phorbol 12-myristate 13-acetate (PMA) (Figure 4). Luciferase expression in the HLR-Elk1 cells wa
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
2. Dual-Luciferase Quantitation in the LMax Microplate Luminometer (MaxLine Application Note #39)
3. Luciferase Reporter Gene Cells and Recombinant Luciferase Dilutions Studied with the CLIPR Luciferase Assay Kit and the LMax Microplate Luminometer (MaxLine Application Note #38)
4. Promegas Multiplexed Luciferase Reporter and Cell Viability Assays performed on the PHERAstar
5. Versatile Reporter Vectors for Monitoring Viral Transduction
6. New Reporter Plasmids for Studying Interferon-Stimulated Signal Transduction Pathways
7. Genetic Reporter Systems
8. Genetic Reporter Systems
9. Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment
10. A Microtiter-based Assay for the Determination of ID50s of b-lactamase Inhibitors Employing Reporter Substrates Detected at UV or Visible Wavelengths (MaxLine Application Note #20)
11. A Comparison Study of Lipid Tranfection Reagents in A549, NIH 3T3 and COS-7 Cell Lines
Post Your Comments:
(Date:12/19/2014)... , Dec. 19, 2014 Naurex Inc., ... novel drugs for diseases of the central nervous system, ... and chief executive officer, will present at the 33 ... presentation will take place at 3:00 p.m. PST on ... in San Francisco, Calif. ...
(Date:12/19/2014)... DUBLIN , Dec. 19, 2014 Research ... announced the addition of the "Technology Innovations ... their offering. http://photos.prnewswire.com/prnh/20130307/600769 ... Innovations in Smart Fabrics" offers a detailed assessment ... Key Findings: 1. ...
(Date:12/19/2014)... 18, 2014 Ed Bilsky, Ph.D., vice ... Center for Excellence in the Neurosciences (CEN), and professor ... been selected as a member of the prestigious Dana ... neuroscientist from Maine to receive this honor. , ... entirely by the Dana Foundation. Its mission is ...
(Date:12/19/2014)... The empty capsules market is experiencing growth ... technological innovations in the empty capsules market. The ... growth of the empty capsules market. , Get ... of Report , The report will enrich both ... to gauge the pulse of the market which ...
Breaking Biology Technology:Naurex to Present at 33rd Annual J.P. Morgan Healthcare Conference 2Technology Innovations in Smart Fabrics (Technical Insights) 2UNE's Ed Bilsky Named Member of Dana Alliance for Brain Initiatives 2UNE's Ed Bilsky Named Member of Dana Alliance for Brain Initiatives 3Empty Capsules Market Hit to Reach $1,820 Million by 2019 - New Research Report by MarketsandMarkets 2Empty Capsules Market Hit to Reach $1,820 Million by 2019 - New Research Report by MarketsandMarkets 3Empty Capsules Market Hit to Reach $1,820 Million by 2019 - New Research Report by MarketsandMarkets 4
... matrix for dry state room temperature transport and ... ... GenVault Corporation, the,leader in dry, room temperature biosample management, ... DNA is a,proprietary inorganic matrix with built-in oxidative protection and,antimicrobial ...
... Pharmasset, Inc.,(Nasdaq: VRUS ) management will present ... from October 13-14, 2008 at the Waldorf Astoria,in ... Officer, will,provide an overview of the company on ... About Pharmasset, Pharmasset is a clinical-stage pharmaceutical ...
... Colo., Oct. 8 GeneThera, Inc.,(Pink Sheets: GTHR), ... Chairman and CEO, Dr. Tony Milici, is featured ... The interview gives viewers an overview of ... press release., To view the clip ...
Cached Biology Technology:GenVault Corporation Launches GenTegra(TM) DNA 2GenVault Corporation Launches GenTegra(TM) DNA 3[video] Dr. Tony Milici, Chairman and CEO of GeneThera, Inc., Discusses Worldwide Rights to E.Coli Vaccine on WallSt.net's 3-Minute Press Show 2
(Date:12/19/2014)... , Dec. 18, 2014   LaunchKey , the ... the post-password and Internet of Things era, today announced ... The venture round was led by Metamorphic Ventures with ... Venture Partners, VegasTechFund, and others.  LaunchKey has raised $4 ... to expand its team and bring LaunchKey to market ...
(Date:12/17/2014)... 2014  HITLAB SM announced today its ... to confirm its adherence to current U.S. Food ... HITLAB to conduct regulated smart device and smart ... patient safety and research quality. "HITLAB ... and delivery with innovative technology," said Laura ...
(Date:12/17/2014)... 2014  Automation is fundamentally transforming the travel ... at international borders. Over the past decade, ePassports, ... veteran travelers to self process through border control ... at an increasing number of airports, seaports, and ... According to Maxine Most , Principal at ...
Breaking Biology News(10 mins):LaunchKey Raises $3 Million in Additional Funding Led by Metamorphic Ventures 2LaunchKey Raises $3 Million in Additional Funding Led by Metamorphic Ventures 3HITLAB Completes GCP Audit for FDA Smart Device & App Compliance 2Automated Border Control (ABC) Transforms the Global Travel Experience With More Than 2500 ABC eGates and APC Kiosks Deployed In Airports, Seaports, and Land Borders Worldwide 2
... at Brown University and Women & Infants Hospital have invented ... potentially powerful new means for conducting fertility research and could ... already used the lab-grown organ to mature human eggs. ... this is the first time that anyone has created a ...
... attack in a young athlete on the playing field is ... more is understood about hypertrophic cardiomyopathy (HCM), a genetic disorder ... young people but which affects people of all ages. So ... by Thomas L. Force, M.D., James C. Wilson Professor of ...
... D.C. (September 14, 2010) -- Piezoelectric materials have fantastic ... And vice-versa, they contract or expand when jolted with ... Greek word meaning to squeeze or press, the piezoelectric ... in several crystals in 1880. But in 1917, a ...
Cached Biology News:Researchers build 'artificial ovary' to develop oocytes into mature human eggs 2Researchers build 'artificial ovary' to develop oocytes into mature human eggs 3Researchers nationwide ask for new focus on 'sudden death' heart disorder 2Researchers nationwide ask for new focus on 'sudden death' heart disorder 3Lead-free piezoelectric materials of the future 2
Complete cell culture media with cytokines...
Consists of 4 separate 100mM solutions (ATP,GTP,CTP and UTP pH 7.5) as sodium salts. 98% Pure by HPLC...
Tracker Datamatrix TRxD reads any 1D and 2D barcodes within 0.5 seconds....
8 different frozen tissues are mounted on positively charged glass slides...
Biology Products: