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:7/30/2014)... (PRWEB) July 30, 2014 Vala ... the U.S. Environmental Protection Agency (EPA). This new ... developmental toxicity models, including tests designed to assess ... and the formation of connections between neurons (synapses), ... neurodifferentiation and synapse formation by environmental pollutants likely ...
(Date:7/30/2014)... Immunology Summit-2014 will ... Investigators, scientists in the field, which includes prominent ... Case Western Reserve University School of Medicine, USA, ... “The effects of recombinant human IL-6 and ... Arya Biragyn, a Senior investigator at National Institute ...
(Date:7/29/2014)... MA (PRWEB) July 30, 2014 ... (UC) and collaboration company, is proud to announce ... account management positions in their Boston, Charleston and ... response to the increased demand for hosted solutions, ... operational efficiency and reduce cost. , SoundConnect was ...
(Date:7/29/2014)... hurdled a number of significant technological challenges in ... technology originally created for the James Webb Space ... Harvey Moseley, a scientist at NASA,s Goddard Space ... electrostatically actuated microshutter arrays that is, those ... as functional as the current technology,s magnetically activated ...
Breaking Biology Technology:U.S. Environmental Protection Agency (EPA) Awards Vala Sciences a Second ToxCast Contract 2U.S. Environmental Protection Agency (EPA) Awards Vala Sciences a Second ToxCast Contract 3Unveiling Breakthroughs in Clinical & Cellular Immunology 2Unveiling Breakthroughs in Clinical & Cellular Immunology 3SoundConnect Organization and Culture is Evolving 2Revolutionary microshutter technology hurdles significant challenges 2Revolutionary microshutter technology hurdles significant challenges 3Revolutionary microshutter technology hurdles significant challenges 4
... PRINCETON, N.J., TOKYO, and REDWOOD CITY, Calif., March ... Inc. (Nasdaq: PDLI ) today announced the ... has acquired PDL,s,rights to IV Busulfex(R) (busulfan), including ... million plus inventory,value., IV Busulfex is an ...
... Calif., March 10 Cepheid (Nasdaq:,CPHD) will present ... March 18 at 10:15 a.m. ET at The ... Chief Executive Officer, will provide a corporate overview., ... may be,accessed on Cepheid,s Web site at ...
... late 2007 ... ... prescribing information for PROCRIT(R) (Epoetin alfa),following guidance from the U.S. Food and ... class., Modifications to the label were based on an unplanned interim ...
Cached Biology Technology:Otsuka Pharmaceutical and PDL BioPharma Close Sale of IV Busulfex 2Otsuka Pharmaceutical and PDL BioPharma Close Sale of IV Busulfex 3Otsuka Pharmaceutical and PDL BioPharma Close Sale of IV Busulfex 4Cepheid to Present at Cowen and Company's 28th Annual Healthcare Conference 2Ortho Biotech Modifies Prescribing Information for PROCRIT(R) (Epoetin alfa) 2Ortho Biotech Modifies Prescribing Information for PROCRIT(R) (Epoetin alfa) 3Ortho Biotech Modifies Prescribing Information for PROCRIT(R) (Epoetin alfa) 4Ortho Biotech Modifies Prescribing Information for PROCRIT(R) (Epoetin alfa) 5Ortho Biotech Modifies Prescribing Information for PROCRIT(R) (Epoetin alfa) 6Ortho Biotech Modifies Prescribing Information for PROCRIT(R) (Epoetin alfa) 7
(Date:7/30/2014)... better decisions about what they eat, the Food and ... to the Nutrition Facts label found on nearly every ... Chemical & Engineering News (C&EN), the weekly news ... updates and the fight that has ensued. , ... that while the Nutrition Facts label has remained largely ...
(Date:7/30/2014)... is a subject of major interest in neuroscience. ... during the distinct stages of Wallerian degeneration: transcription ... dedifferentiation. Although gene expression responses in the distal ... injury are known, differences in gene expression between ... Dengbing Yao and co-workers from Nantong University, China ...
(Date:7/30/2014)... JOSE, Calif. , July 30, 2014 /PRNewswire/ ... partnership with the United States Air Force Research ... proposed by Binghamton University ... human performance monitoring (HPM). The $425,000 project, with ... electronics packaging firm i3 Electronics, Inc. ( ...
Breaking Biology News(10 mins):Nano-Bio Manufacturing Consortium Awards Funding to Binghamton University for Development of Electronics and Biometric Sensor Platforms 2Nano-Bio Manufacturing Consortium Awards Funding to Binghamton University for Development of Electronics and Biometric Sensor Platforms 3
... could see humble cotton thread emerge as a core ... such as kidney failure and diabetes. In a ... and sewing needles to literally stitch,together the uniquely low-cost ... postage stamp. Microfluidic analytical devices, which have been ...
... University of Wisconsin-Madison engineers has developed a highly ... a biomass derivative, into the chemical equivalent of ... 95 percent of the energy from the original ... dioxide under high pressure for future beneficial use. ...
... new University of Colorado at Boulder study shows mice ... with a deadly and poorly understood human inflammatory disease, ... The human disease, known as Hemophagocytic Lymphohistiocytosis, or HLH, ... percent and 90 percent of its victims, said Diane ...
Cached Biology News:How long is a piece of thread? Long enough to save a life 2New process yields high-energy-density, plant-based transportation fuel 2New University of Colorado paper shows novel way to study human inflammatory disease 2
... pLivSelect is a ... for recombinant identification. Recombinant ... colony survival, eliminating the ... screening. This quick and ...
... Signal Enhancer does for enzyme-/substrate-based blotting what intensifying ... the signal up to 10-fold (or one order ... Western Blot Signal Enhancer membrane treatment is a ... to your current Western blotting protocol. The result ...
... multi-functional program that searches for ... sequence file for polymerase chain ... mutagenesis, and various hybridization applications. ... secondary structure of oligonucleotides based ...
...
Biology Products: