Navigation Links
Performance of Ad-A-Gene EGFP-Glucocorticoid Receptor (EGFP-GCCR): an adenoviral vector gene delivery system

) ratio is the population-averaged ratio of sampled nuclear and cytoplasmic intensities measured in the EGFP signal channel.

* Time-controlled dispensing of compound solutions to the cells can be avoided by completing the assay using live cells and then fixing the cells prior to imaging. Cells should be fixed at the peak translocation time point. Fixing of cells reduces the handling requirements for post-assay material to BSL-1 categorization.


Assay protocol (kinetic live cell)
Test compounds (e.g., 200-nM dexamethasone containing 2.5-μM Hoechst nuclear dye) were added at 100-μl/well to cells in CS-FCS medium (100 μl) that had been transduced 24 h earlier as described previously. Plates were incubated for 30–60 min at 37 ºC, 5% CO2.

Cells were imaged on an appropriate detection instrument, using the image acquisition method as described above for the fixed-cell assay protocol. Analysis of images also followed the method outlined above for the fixed-cell assay protocol.

Note: at all stages of the live-cell assay, the assay plate must be contained within BSL-2 level facilities.


Results
Viral transduction efficiency measurement from flow cytometry data
The primary step to setting up a functional cellular assay involving adenoviral transduction is to determine the optimal MOI by titrating into target cells. We have made use of flow cytometry to quantitate the EGFP signal from cells transduced with the EGFP-GCCR adenoviral vector. Analysis of data allowed calculation of the transduction efficiency in HeLa cells at three MOI loadings (Table 1). HeLa cells were set up at 1 × 106 cells/well of a 6-well plate and virus was added at 0 (control), 25, 50, and 100 MOI. The plates were stored for 24 h and then the cells were trypsinized and resuspended in PBS followed by gated analysis on a FACSCalib
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8 9 10

Related biology technology :

1. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
2. Comparing Fidelity and Performance of Proofreading PCR Enzymes
3. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases
4. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
5. Performance Comparisons of Commercial RT-PCR Systems
6. Restriction Endonucleases Undergo Rigorous QC Testing to Ensure Optimal Performance
7. Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance
8. Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance
9. Challenge the Performance of Your Hot-Start PCRs with FastStart Taq DNA Polymerase and the Novel FastStart High Fidelity PCR System
10. Highly Sensitive microRNA Array Performance
11. High Throughput RNA Isolation for High Performance Microarray Analysis
Post Your Comments:
(Date:3/26/2015)... , March 26, 2015 PAREXEL ... global clinical research organization , announced ... definitive agreement to acquire all of the ... a leading provider of specialized pharmacovigilance services, ... the collection, detection, assessment, monitoring, and prevention ...
(Date:3/26/2015)... America," the popular news television program that airs in numerous ... film actor James Earl Jones, has announced that it will ... , Botany is commonly described as the scientific study of ... more complicated than that. Scientists and other professionals study a ... smallest bacteria known to man to the largest trees that ...
(Date:3/26/2015)... WriteResult, a premier provider of ePRO ... University’s School of Public Health and New Haven Farms ... 4 month-long project. The goal of the study was ... with free access to fresh produce and nutrition education ... like high blood pressure, diabetes and weight gain. Patients ...
(Date:3/25/2015)... (PRWEB) March 26, 2015 Agnition announced ... episode of American Farmer, airing for the first time ... segment of American Farmer will explore the extremely important ... crop producers with a patented Microbial Catalyst™ technology that ... greater yields. The segment will air again on April ...
Breaking Biology Technology:PAREXEL Announces Execution of Definitive Agreement to Acquire Quantum Solutions India, Strengthening Leadership In Pharmacovigilance Services 2PAREXEL Announces Execution of Definitive Agreement to Acquire Quantum Solutions India, Strengthening Leadership In Pharmacovigilance Services 3PAREXEL Announces Execution of Definitive Agreement to Acquire Quantum Solutions India, Strengthening Leadership In Pharmacovigilance Services 4Yale and New Haven Farms Improving Patient Health with WriteResult's myPROpad™ 2Yale and New Haven Farms Improving Patient Health with WriteResult's myPROpad™ 3Agnition to be Featured In Upcoming Episode of American Farmer on RFD-TV 2
... , , ... , ... for culturing cells in a BioFlo ,110 benchtop fermentor/bioreactor system. ... and control modules to,enable growth of a wide variety of cultures, ...
... Microscale on-membrane PMF analysis can be ... function of a chemical printer (Refer to AXIMA Application 14, 17). ... chemical printer enables MS analysis with high mass accuracy ( , ... separated by 2-dimensional electrophoresis were transferred to a PVDF membrane for ...
... Achieving Ultra Fast Liquid Chromatography ... , Prominence UFLC has ... this respect, we are successful,in demonstrating high speed and separation performance,without ... with other fast,LC systems., Smaller ...
Cached Biology Technology:CHO Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor 2CHO Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor 3CHO Cell Culture Application Using a BioFlo 110 Benchtop Bioreactor 4Direct On-Membrane MS/MS Analysis using the Chemical Printer (CHIP-1000) and AXIMA-QIT 2Prominence UFLC 2Prominence UFLC 3Prominence UFLC 4Prominence UFLC 5Prominence UFLC 6Prominence UFLC 7Prominence UFLC 8
(Date:3/20/2015)... Mar. 20, 2015 Research and Markets ... of the "India Sensors Market Forecast and ... The sensor market is projected to grow ... Consumer electronics, automotive, industrial and healthcare sectors ... the country. In addition, adoption of MEMS technology ...
(Date:3/20/2015)... and Markets ( http://www.researchandmarkets.com/research/vkzvcs/iris_biometrics ) has announced the ... Report" report to their offering. This report ... The report provides separate comprehensive analytics for the US, ... Europe , Asia-Pacific , ... . Annual estimates and forecasts are provided ...
(Date:3/17/2015)... Minn. , March 17, 2015 ... technology company specializing in clinical study management systems, ... be in attendance at ARENA,s Outsourcing in Clinical ... NC.  This event provides an excellent forum for ... easy-to-use, flexible and affordable software-as-a-service (SaaS) eClinical technology ...
Breaking Biology News(10 mins):India Sensors Market Forecast and Opportunities 2020 - STMicroelectronics held the largest share in 2014 2India Sensors Market Forecast and Opportunities 2020 - STMicroelectronics held the largest share in 2014 3Iris Biometrics - Global Strategic Business Report 2015: System & Network Access Control - A Robust Market Opportunity in the Offing 2Iris Biometrics - Global Strategic Business Report 2015: System & Network Access Control - A Robust Market Opportunity in the Offing 3Iris Biometrics - Global Strategic Business Report 2015: System & Network Access Control - A Robust Market Opportunity in the Offing 4Iris Biometrics - Global Strategic Business Report 2015: System & Network Access Control - A Robust Market Opportunity in the Offing 5Connect With MedNet Solutions At Outsourcing In Clinical Trials Southeast 2
... the event of a standoff between the United States ... elected president, know enough about the physics of nuclear ... nation toward reduced greenhouse gas emissions, would John McCain ... carbon footprint? If not, University of California, Berkeley, ...
... the California Institute of Technology have developed a ... drugs, including antiplaque toothpaste additives, antibiotics, nicotine, and ... paper describing the research, now available online, will ... September issue of Nature Chemical Biology . ...
... the Smithsonian Institution have discovered a new species of ... to the scientific community. Their findings were published in ... The newly found olive-backed forest robin (Stiphrornis pyrrholaemus) was ... and rump. Adult birds measure 4.5 inches in length ...
Cached Biology News:New book tutors future presidents and public on science behind the headlines 2New book tutors future presidents and public on science behind the headlines 3New book tutors future presidents and public on science behind the headlines 4New book tutors future presidents and public on science behind the headlines 5Caltech engineers build mini drug-producing biofactories in yeast 2Smithsonian scientists discover new bird species 2Smithsonian scientists discover new bird species 3