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

porter cell lines that express the GAL4-CREB, GAL4-Elk1, or GAL4-cJun. In the HLR-CREB, HLR-Elk1, and HLR-cJun cell lines, luciferase activity reflects the activation status of the chimeric trans-activator protein and, therefore, the respective signaling pathways. By performing simple luciferase assays, researchers can directly evaluate the effects of extracellular stimuli on signaling pathways converging at CREB, Elk1, or c-Jun. Use of these cell lines eliminates transfections, thereby saving time while avoiding the variables that can be caused by fluctuations in transfection efficiencies. The HLR cell line can be used to assess protein-protein interactions in mammalian cells,11 while the HLR-CREB, HLR-Elk1, and HLR-cJun cell lines can be used to study transcription mechanisms. Another application for these cells includes adaptation for high-throughput drug screenings, since many components of signal transduction pathways are potential targets for drug development efforts.

Establishing Stable Reporter Cell Lines

Fig.1

To create stable reporter cell lines with the PathDetect trans-reporting systems, we tried several approaches. Figure 1 outlines our most successful protocol: We first transfected the pFR-Luc plasmid with a hygromycin expression cassette into HeLa cells. Through successive rounds of selection in antibiotic medium, hygromycin-resistant clones were isolated and tested with the PathDetect CREB, Elk1, and c-Jun trans-reporting systems. Cell lysates from these stable clones were then prepared, assayed for luciferase activity, and analyzed. One clonal line, designated as HLR for the HeLa luciferase reporter cell line, was chosen for further experimentation. It gave the highest levels of luciferase activity when tested with the trans-reporting sys
'"/>

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:7/30/2015)... July 30, 2015 AACC welcomed thousands ... 2015 AACC Annual Meeting & Clinical Lab Expo ... 26–30. The meeting showcased revolutionary advancements in clinical ... of healthcare providers to diagnose patients quickly and ... treatment. As of Wednesday, July ...
(Date:7/30/2015)... ... July 30, 2015 , ... ... Francois Vieillard as Sales & Marketing Director. , With more than 25 ... be responsible for reinforcing Tronics’ business development activities worldwide. He brings to the ...
(Date:7/30/2015)... July 30, 2015 According to ... (Software, Molecular Modeling, PB/PK, PKPD, Trial Design, Toxicity Prediction, ... Preclinical, Clinical Trial), by End Users - Global Forecast ... expected to reach USD 2,107.99 million by 2020 from ... 15.29%. Browse more than ...
(Date:7/30/2015)... 30, 2015   Aratana Therapeutics, Inc . ... focused on the licensing, development and commercialization of ... that its strategic partner VetStem Biopharma, Inc. has ... of AT-016, an adipose-derived allogeneic stem cell product ... osteoarthritis pain in dogs. The double-blinded, ...
Breaking Biology Technology:Innovative Research on Cancer, Heart Disease, and Infectious Diseases Draws More than 17,500 Attendees to 2015 AACC Annual Meeting 2Innovative Research on Cancer, Heart Disease, and Infectious Diseases Draws More than 17,500 Attendees to 2015 AACC Annual Meeting 3Innovative Research on Cancer, Heart Disease, and Infectious Diseases Draws More than 17,500 Attendees to 2015 AACC Annual Meeting 4Tronics appoints Francois Vieillard as Sales and Marketing Director 2Biosimulation Market Worth 2,107.99 Million USD by 2020 2Biosimulation Market Worth 2,107.99 Million USD by 2020 3Biosimulation Market Worth 2,107.99 Million USD by 2020 4Aratana Therapeutics Announces Positive Pilot Field Study for AT-016 2Aratana Therapeutics Announces Positive Pilot Field Study for AT-016 3Aratana Therapeutics Announces Positive Pilot Field Study for AT-016 4Aratana Therapeutics Announces Positive Pilot Field Study for AT-016 5
... , May 13 /PRNewswire-Asia/ -- A signing ceremony for ... (TJAB) and Genzyme Corporation was held in the,Tianjin Economic-Technological Development Area ... , , ... Zihe, President of TJAB and,President of Nankai University, He Shushan, Director ...
... TRIANGLE PARK, N.C. , May 13 ... today announced the commencement of a first-in-human study of ... with in vitro activity against both tenofovir-sensitive ... potential to increase efficacy and decrease toxicity as compared ...
... Repligen Corporation (Nasdaq: RGEN ) announced today ... with the Food and Drug Administration (FDA) for a Phase ... inhibitor.  This is a double-blind, single ascending dose, Phase 1 ... profile of RG2833 in up to 40 subjects.  This study ...
Cached Biology Technology:Tianjin International Joint Academy of Biotechnology and Medicine, Genzyme Enter into Strategic Partnership 2Chimerix Initiates Phase 1 Study of CMX157 2Chimerix Initiates Phase 1 Study of CMX157 3Repligen Files Investigational New Drug Application with FDA for First Drug Targeting the Core Genetic Defect of Friedreich's Ataxia 2Repligen Files Investigational New Drug Application with FDA for First Drug Targeting the Core Genetic Defect of Friedreich's Ataxia 3Repligen Files Investigational New Drug Application with FDA for First Drug Targeting the Core Genetic Defect of Friedreich's Ataxia 4
(Date:7/31/2015)... , 31 de julio de 2015 ... (ICG-10,  www.icg-10.org ) del 22 al 25 de octubre ... La conferencia celebra su décimo aniversario este ... convertido en una de las reuniones anuales más influyentes ... es una de las conferencias científicas más dinámicas, entusiastas ...
(Date:7/31/2015)... 2015 La 10e Conférence internationale sur ... par le BGI du 22 au 25 octobre ... Chine. La conférence célèbre son 10e ... l,ICG est devenue l,une des réunions annuelles mondiales ... et c,est aussi la plus dynamique, la plus ...
(Date:7/31/2015)... China , 31. Juli 2015 Die 10. ... wird von BGI veranstaltet und findet vom 22. ... China , statt. Die ... Seit ihrem Start 2006 ist die ICG weltweit ... „omik" geworden. Sie ist eines der dynamischsten, enthusiastischsten ...
Breaking Biology News(10 mins):La 10ª Conferencia Internacional sobre Genómica (ICG-10) comienza en octubre 2La 10ª Conferencia Internacional sobre Genómica (ICG-10) comienza en octubre 3La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 2La 10e International Conference on Genomics (ICG-10) doit s'ouvrir en octobre 3Die 10. internationale Konferenz zu Genomik (ICG-10) beginnt im Oktober 2Die 10. internationale Konferenz zu Genomik (ICG-10) beginnt im Oktober 3
... straight to the uncovered food in your kitchen? Now ... over proportionally huge distances, using just very poor eyesight ... have significant benefits in the development of autonomous robots ... processes. , Scientists at the University of Sussex, ...
... the same metabolic disorders that have led to an epidemic ... of research conducted at Penn State University that are due ... the Proceedings of the National Academy of Science and also ... on 21 November. The discovery expands the known taxonomic ...
... heart failure and other health conditions, blood draws and diagnostic ... though, chemical or physiologic changes silently cause damage that is ... the future a tiny device, one the size of a ... to monitor and detect abnormalities, and could then relay data ...
Cached Biology News:How ants find their way 2Dragonfly's metabolic disease provides clues about human obesity 2Dragonfly's metabolic disease provides clues about human obesity 3Dragonfly's metabolic disease provides clues about human obesity 4Dragonfly's metabolic disease provides clues about human obesity 5Cardiologist's 'living chip' changes science of disease monitoring 2Cardiologist's 'living chip' changes science of disease monitoring 3Cardiologist's 'living chip' changes science of disease monitoring 4
...
... Although the mixed TAMRA isomers are ... isomers are increasingly preferred for labeling peptides ... in HPLC purification that is often required ... often used than 6-TAMRA for labeling peptides ...
Rabbit polyclonal to GPCR G2A ( Abpromise for all tested applications). entrezGeneID: 29933 SwissProtID: Q9UNW8...
Alkali soluble casein is the recommended blot blocking reagent for use with the HisTag Monoclonal Antibody (Novagen) on Western blots....
Biology Products: