Navigation Links
Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells

evels of expression of exogenous proteins typical of transiently transfected cells. It was therefore necessary to optimize our transfection experiments in order to produce enough positively transfected cells for data analysis. In an attempt to overcome these difficulties, we compared the effectiveness of two liposome-based transfection reagents in SW13 cells using pEGFP-C3 as a control vector.

Transient Transfection of the pEGFP-C3 Vector into SW13 Cells

SW13 cells (8 x 104) were plated in a 24-well culture dish and allowed to attach overnight so that the cell density was between 60% and 80% confluent. In order to compare the relative efficiencies of the two liposome-mediated reagents, we used the manufacturers recommendations for concentration of reagent and DNA as a starting point for our experiments. Additional transfections were performed that used more or less of either the reagent or DNA. All procedures were performed according to the manufacturers recommendations, including the use of serum-free media for LipofectAMINE reagent-mediated transfections and serum-enriched media when adding LipoTAXI transfection reagent and DNA solutions to SW13 cells. Cells were exposed to the DNA-liposome complex for 5 hours at 37C in a humidified incubator with 5% CO2. Serum-containing medium was added to the cells, and the cells were incubated overnight. Cells were then fixed with 4% paraformaldehyde at room temperature for 25 minutes and rinsed with deionized, distilled water. Coverslips with attached cells were mounted in glycerol containing 2.5% 1,4-diazabicyclo(2.2.2)octane (DABCO) then viewed and photographed using a Nikon Microphot FX light microscope. Transfection efficiency was determined by counting fluorescent and total cells from six random fields for each condition.

Figure 1 shows fluorescent images of
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell Line
2. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
3. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
4. Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance
5. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
6. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
7. High-Efficiency Transfections Achieved with New Low-Toxicity Reagent
8. Efficient Transfection of Neurospora Crassa
9. Eppendorf Multiporator Transfection Protocols for Eukaryotic Cells
10. Efficient Transfection of Neurospora Crassa
11. Eppendorf Multiporator Transfection Protocols for Eukaryotic Cells
Post Your Comments:
(Date:10/22/2014)... October 21, 2014 The North American ... market North America with analysis and forecast of revenue. ... and is expected to reach $623.6 million by 2018, ... , Browse through the TOC of the North American ... the in-depth analysis provided. This also provides a glimpse ...
(Date:10/22/2014)... Involution Studios , a healthcare software ... infographic, Understanding Ebola . The goal of ... to follow informative tool for anyone wanting to know ... prevention. , "As the news has spread in all ... represents not only a healthcare crisis with global impact, ...
(Date:10/22/2014)... October 22, 2014 Currently offering ... A & D Weighing, Rice Lake, and Ohaus, ... Balance . The Sartorius CPA Semi-Micro Balance is ... high-quality, precise, and user-friendly laboratory balance. , Sartorius ... equipment, their laboratory balances are well known for ...
(Date:10/22/2014)... Hill, SC (PRWEB) October 22, 2014 ... expansion of its cardiovascular pharmacogenetics menu, which enables ... patient outcomes. With PCLS’s evidence-based results, healthcare ... needs and optimize their therapy, while minimizing risks ... In the U.S., according to the FDA [1] ...
Breaking Biology Technology:The North American crystal oscillator market is expected to reach $623.6 million by 2018 - New Report by MicroMarket Monitor 2The North American crystal oscillator market is expected to reach $623.6 million by 2018 - New Report by MicroMarket Monitor 3The North American crystal oscillator market is expected to reach $623.6 million by 2018 - New Report by MicroMarket Monitor 4Involution Studios' Ebola Infographic Provides Key Disease Information and Statistics at a Glance 2Pipette.com Launches the Addition of Sartorius CPA Semi-Micro Balance to Their Product Portfolio with Special Promotional Pricing 2Pipette.com Launches the Addition of Sartorius CPA Semi-Micro Balance to Their Product Portfolio with Special Promotional Pricing 3PCLS Announces Expansion of Cardiovascular Pharmacogenetic Testing Menu 2PCLS Announces Expansion of Cardiovascular Pharmacogenetic Testing Menu 3
... 16 Medical and technical experts at Wyle are ... Station. Their efforts will help astronauts cope with effects ... in a future mission to Mars. , "If successful, ... exploration missions and provide us with better insight into ...
... , , SEATTLE, Nov. 16 ... "Company") today announced that the Company,s product candidate PX-866, ... Information, Inc., as one of the Top 10 most ... partnering. , The selection was made by an independent ...
... , LONDON, Nov. 16 The ... field of biomarker research will test the frontiers of biomedical ... day and this rapid development promises excellent growth potential for ... in biomarker validation and assay development, there is a huge ...
Cached Biology Technology:Wyle Personnel Take Part in Important Studies Aboard the International Space Station 2Wyle Personnel Take Part in Important Studies Aboard the International Space Station 3Oncothyreon's PX-866 PI-3 kinase inhibitor named among Top 10 most promising oncology products for partnering by Windhover 2Oncothyreon's PX-866 PI-3 kinase inhibitor named among Top 10 most promising oncology products for partnering by Windhover 3Frost & Sullivan Reveals Strategies on Turning Challenges into Opportunities in the European Biomarkers Market 2
(Date:10/19/2014)... access in India over the last 30 years ... emissions growth during that time, according to a ... Climate Change ., "Energy access is fundamental to ... life, including education, communication, and health," says IIASA ... While increased energy access is widely agreed to ...
(Date:10/17/2014)... critical and pertinent for practicing physicians and clinicians given ... The Journal, Disaster Medicine and Public Health Preparedness ... Public Health, to surround the public, medical professionals and ... , On October 17, the journal published ... was prepared by Dr. Eric Toner, internist and emergency ...
(Date:10/16/2014)... the human body. Battles are won, lost or ... this stalemate—known as tumor dormancy—can last up to ... that is poorly understood. , A new ... Torquato, a Professor of Chemistry at Princeton University, ... and the switch to a malignant state. Published ...
Breaking Biology News(10 mins):Improved electricity access has little impact on climate change 2Modeling tumor dormancy 2
... few things as important on, and especially off, Earth as ... don,t have seconds to spare, which is why development of ... monitor can measure the concentrations of multiple gases within the ... designed the Multi-Gas Monitor to be resistant to shock and ...
... the right shape might be just as important in a ... an important role in the effectiveness of cells grown to ... design new structures that enable cells to "shape up," researchers ... come up with a way to measure, and more importantly, ...
... sound very durable. But the new type of micro-factories invented ... They will actually be placed inside a patient,s body ... will gradually dissolve and disappear in the patient,s urine once ... that Associate Professor of Chemistry Alexander Zelikin, Aarhus University, has ...
Cached Biology News:Rugged, rapid monitor safeguards space crews 2Rugged, rapid monitor safeguards space crews 3Shape-sifting: NIST categorizes bio scaffolds by characteristic cell shapes 2Tiny soft medicine factories 2
Rabbit polyclonal to ErbB 2 (phospho Y1248) ( Abpromise for all tested applications). entrezGeneID: 2064 SwissProtID: P04626...
The Microseal 384 plate positioner ensures precise positioning of Microseal 384-well plates in the x-, y-, and z-dimensions, for accurate automated liquid handling....
... sealing material is suitable for PCR ... down heated lid), especially 384-well formats ... for sample retrieval. Multiple seals can ... Ideal for compound management and high ...
... Nikon's Digital Sight DS-U2 camera controller is ... is specifically designed to interface any of the ... a PC computer through a USB2.0 interface. The ... complicated and cumbersome installation by equipping both hub ...
Biology Products: