toxicity

New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter

...rGFP expression. Most importantly, the low toxicity of hrGFP expression ensures that use of Vitality vectors results in artifact-free transient and stable experiments. REFERENCES 1. Felts K., Rogers, B., Chen, K., Ji , H., Sorge, J., and Vaillancou...

Reliable Assessment of Micronuclei for Genotoxicity Testing in Mice or Rats

...measure stem cell toxicity. No substantial toxicity was evident at any of the doses tested. Easy and Reproducible Procedure The flow cytometric procedure employed in the kit is used routinely at LitronLaboratories with excellent results. While the procedure critically depends onfixing the cells reli...

A Simple and Rapid Technique to Purify High-Quality Midiprep DNA

...fficult to separate fromplasmid DNA and results in toxicity to cultured mammalian cells. 2 TheStrataPrep EF midiprep protocol provides an optional extraction of endotoxins,which are reduced to 0.1 to 0.2 EU/g DNA (endotoxin units per microgram ofplasmid DNA); hence, high-efficiency transfection results. Fas...

Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells

...ions than LipofectAMINE reagent. Becauseof the low toxicity and increased efficiency of LipoTAXI transfection reagent,it is an invaluable tool for research that is dependent on a higher number oftransfected cells in the total cell population. REFERENCES Sarria, A.J., et al. (1990) J. Cell Biol. 111:...

A Comparison Study of Lipid Tranfection Reagents in A549, NIH 3T3 and COS-7 Cell Lines

...nstant. For the other transfection agents, toxicity to cells increased as their amounts increased. ...r comparison shows the superior efficiency and low toxicity of LipoTAXIreagent, features that make it the best transfection reagent for transfectionsin A549, CO...

Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance

..., thesestudies confirm the high efficiency and low toxicity of LipoTAXI transfectionreagent. Using transfecti...gave thehighest transfection efficiency and lowest toxicity in all three cell lines. Transfection Efficiencies: Effect of Mycoplasma Contamination When standa...

Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells

...s a novel liposome reagent that offerslow cellular toxicity and high transfection efficiencies for adherent an...r transfection efficiencies and increasing toxicity is shown in figures 1 and 2. In these experiments, 1 x 105 PrEC cells were plated in 12-well...

Simple and Rapid Isolation of Plant RNA Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit

...vents such as phenol or chloroform; however, their toxicity and the resulting cumbersome disposal may make alternative protocols more desirable. This application protocol demonstrates that the Eppendorf Perfect RNA Maxi Kit for eukaryotic cells is ideally suited for the isolation of to...

Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent

...g -galactosidase (-gal). No obvious signs of toxicity were detected after transfection. The combination ...-field images, there are no obvious signs of toxicity in the transfected cells. GFP fluorescence is very bright throughout the transfected neurons a...

Transfection Reagent Selector Kit Handbook

...aluation of gene expression - is any toxicity apparent? Check and note the incubation time before harvesting for evaluation of gene expression. To assay for gene expression, proceed as follows: For tran...

Analyzing Trace-Level Impurities of a Pharmaceutical Intermediate Using an LCQ Deca Ion Trap Mass Spectrometer and the Mass Frontier Software Package

...ion, distribution, metabolism, excretion and toxicity of compounds, their metabolites and impurities must be assessed quickly and accurately to characterize potential lead compounds. High-throughput analysis of impurities is particularly challenging at the low sample conce...

Cell Proliferation and Viability Measurement

... compounds in toxicology testing,and cell-mediated toxicity can be assessed whenquantifying cell growth. The first step in any experiment is the decision whetherto test for cell proliferation or viability parameters,depending on the objective of a study. These parametersare measured by assaying f...

Flow cytometric analysis of human primary cells using the Agilent 2100 bioanalyzer and on-chip staining

... live transfected cells it is importantto take the toxicity of the transfection reagent into account.For this experiment, a ratio of 1:6was found to be optimal. In thosecases where enough cells wereobtained, samples were measuredin parallel on a flow cytometer(figure 1D - only GFP histogramsare shown). We als...

siRNA transfection optimization with the Agilent 2100 bioanalyzer

...easingamounts due to too low siRNAconcentration or toxicity when inexcess. The Transfection Viability withRNAiFect was evaluated assuggested by QIAGEN protocolsfor Hela cells, in the range from0.5 to 1.5 g siRNA/sample in a 24well plate and siRNA to RNAiFectratios from 1:3 to 1:9. It shows veryhigh Tra...

Advancing the quality control methodology to assess isolated total RNA and generated fragmented cRNA

...rvices related togene activity in human diseaseand toxicity that enable globalpharmaceutical and biotechnologycompanies to optimize the time,risk and cost of drug discoveryand drug development. In order todo this, the quality control methodsmust ensure that only thehighest quality samples proceedthrough our h...

Silencer Validated siRNAs

...sample without causing cytotoxicity (e.g., toxicity is usually a problem when transfecting cells with greater than 200 nM siRNA). * Most importantly, using sequences that have been verified to effectively induce gene silencing avoids the need to screen siRNAs, and thus saves ...

A New Way to Assess siRNA Delivery Efficiency

...induced knockdown AND transfection-induced toxicity Use with Silencer GAPDH siRNA Controls a... levels of gene silencing while minimizing toxicity associated with the transfection process. Ambions Silencer GAPDH siRNA is an ide...

Choose the Right Transfection Agent for Your RNAi Experiment

...on of polyamines, the reagent exhibits low toxicity and can be used in the presence or absence of serum. siPORT XP-1 is simple to use -- reagent/DNA complexes can be added directly to growing cells in serum-containing culture media, and the media can be left unchanged until...

Inducing RNAi with siRNA Cocktails Generated by RNase III

...oes not exhibit higher levels of toxicity (data not shown) or nonspecific effects on gene expression as compared to individual chemically synthesized siRNAs. The use of RNase III to prepare siRNA cocktails eliminates the need for expensiv...

Dicer vs. RNase III for Preparation of siRNA Cocktails

...te samples from Panel A were examined for cellular toxicity using a Trypan Blue Exclusion Assay. Background levels of cell death of 2% were recorded in nontransfected cells; none of the transfected samples showed significantly higher levels of cellular toxicity. Figure 3....

Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection

... GAPDH expression in all three cell types. Some toxicity was observed at the highest siRNA dose tested (2.66 g), but there was a complete absence of non-specific effects at 1.33 g siRNA (Figure 3). To extend these results, <A href="http://www.ambion.com/catalog...

Optimizing siRNA Transfection for RNAi

... NeoFX Transfection Agent shows minimal toxicity and yields a broad range of silencing acti...d. In this cell type, siPORT NeoFX shows minimal toxicity and yields a broad range of silencing activity. Some reagents are not as flexible an...

Optimization of Biolistic Transformation Using the Helium-Driven PDS-1000/He System

...able transformants due to the demonstrated toxicity of tungsten in some plant cells (Russell, et al., ...nford, J. C., Physical trauma and tungsten toxicity reduce the efficiency of biolistic transformation, Plant Physiol., 98 , 1050-1056 (1992b). ...

Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cells

...nford, J. C., Physical trauma and tungsten toxicity reduce the efficiency of biolistic transformation, Plant Physiol., 98 , 1050-1056 (1992). 12. Sanford, J. C., Smith, F. D. and Russell, J. A., Optimizing the biolistic process for different biological applications, Meth. Enzym...

The measurement of DNA condensation and mitochondrial changes during apoptosis and necrosis in cell based assays

...633 are also observed. Thus, although little or no toxicity is detected using traditional measurements such as...ell number measurements are that there is no overt toxicity occurring in the system at any dose of diclofenac. By contrast, it is clear from the data acquired w...

Simultaneously characterizing and quantifying chloramphenicol and its metabolite using LC/MS/MS

... toxic effects on humans. The main potential human toxicity is depression of red blood cell production in bone marrow leading to a plastic aneamia and leukemia in children especially. It is important to identify chloramphenicol and monitor its exposure levels in different subjects. Quantification of c...

Speciation of Arsenic by LC-ICP-MS

...ion of these species can greatly assist risk-based toxicity assessments. When Liquid Chromatography (LC) is interfaced with Inductively Coupled Plasma Mass Spectrometry (ICP-MS), species elute one by one from the LC column directly to the ICP-MS for detection by elemental speciation. The coupling of LC...

Quantitation of Digoxin in Human Plasma Using Negative Ion Electrospray Ionization LC/MS

... of Digoxin in human serum is 0.5 - 2.0 ng/mL with toxicity starting at 4.0 ng/mL. Because of Digoxins high potency, doses are administered from 0.125 mg to a maximum of 0.5 mg, once a day. Periodic plasma level measurements are critical for establishing the proper dose administration. The onset of action fro...

Simultaneous Detection of CYP3A4, CYP2D6 and CYP2C9 Metabolites with a Single, Sensitive, LC/MS/MS Method

...g-drug interactions 3 , drug side effect(s) 4 and toxicity (including hepatotoxicity). Also relevant to CYP testing is the discovery that polymorphisms in the genetic sequences expressing these enzymes leads to populations of people whose ability to metabolize certain classes of drugs may be attenuated in co...

A DELFIA Assay for Inhibition of PARP

...erase gene affects oxygen radical and nitric oxide toxicity in islet cells. J. Biol. Chem. 270 , 11176-11180 5. Davidson, P. F. 1959. The effect of hydrodynamic shear on the deoxyribonucleic acid from T2 and T4 bacteriophages. Proc. Natl. Acad. Sci. 45 , 1560-1568 6. Purnell, M. R. and Whi...

measurement of green fluorescent proteins and a red fluorescent protein in live cells in the Analyst GT multimode reader

... The fluorescent proteins are stable, have minimal toxicity and have the ability to generate visible fluorescence in vivo without the need for external cofactors. Thus they can be used as molecular tags or as independent reporters to visualize, track and quantify many different cellular processes, including p...

Ca++-Induced G Protein Coupled Receptor (GPCR) Activation Monitored via Aequorin Luminescence in the LMax Microplate Luminometer (MaxLine Application Note #42)

...ells upon receptor activation. The lack of reagent toxicity and ease of use of aequorin-expressing cells may provide a useful alternative method to calcium-sensitive dye-loading procedures for determining receptor activation. The Lmax offers high assay sensitivity and automated reagent injection in a 96-well ...

Analysis of a Verapamil Microsomal Incubation using Metabolite ID and Mass FrontierTM

...ed to completeness, especially when regulatory and toxicity issues are involved. By the time a drug candidate reaches the latter stages of development, a team of scientists might spend years exhaustively characterizing its metabolic products. LC/MS has become the technique of choice in these metabolism...

Near Infrared Technology and Optical Agents For Molecular Imaging

...stemsand there are questions about their long-term toxicity 4 . A number of studies have been published using NIRdyes. Two of the dyes commonly used for opticalimaging are Cy5.5 and IRDye 800CW. Cy5.5 hasbeen used in the past primarily due to the lack ofother candidate dyes suitable for imaging. Cy5....

The Genesis of Gendicine: The Story Behind the First Gene Therapy

...logy tests? Peng: In our tests, no genetic toxicity was observed. Results from both a Chinese hamster lung (CHL) cell-chromosomeaberration test and mouse bone-marrow micronucleus assay were negative. Gendicine will not replicate in the infectedcells and is incapable o...

Alzheimer's drug developer lands $1.2M in venture funding

...ssion of the disease, an effective drug must block toxicity and pass into the central-nervous system by crossing the "blood-brain barrier." John Neis, founding partner of Venture Investors, said the annual sale of Alzheimer's drugs is $2.3 billion, but while available drugs have benefits for patients, none o...

Biotech firm uses live heart tissue in drug screens

...lated and invalid leads and protein targets due to toxicity issues, for example, at very early stages of the d... growth per year, and the therapeutic efficacy and toxicity testing markets comprise approximately $1.1 billion in the U.S., she said. Annac said that she is a...

Biotech firm has strokes, Alzheimer's in its sights

...ight someday come to the rescue has been used as a toxicity indicator in scientific research for 40 years, but until now it has never been investigated for its therapeutic qualities. To produce it in the necessary quantities for medicine, the aequorin cells must be reproduced, a process that expresses the pro...

Mithridion wins grants for Alzheimer's research

...ssion of the disease, an effective drug must block toxicity and pass into the central-nervous system by crossing the "blood-brain barrier." Value validation To validate the drug leads, Mithridion will treat cells with a variety of drug candidates and measure the resulting gene activation. Later, the compa...

Medical College researcher working to make genetic-based medicine a reality

...varies significantly, and they have a high rate of toxicity and treatment failure. "One of the problems with ...too slow or too well, that could be a predictor of toxicity or treatment failure," he said. Collaboration call As part of the pilot studies, Stephens pla...

Medical College researchers win federal grants

...varies significantly, and they have a high rate of toxicity and treatment failure. By understanding how genetic variation impacts drug transporter function, the medical community may be able to improve, and personalize, treatment strategies that include thiopurines. Hammer to sickle Medical College resear...

Wisconsin to show up at BIO in numbers

...osciences, and Cellular Dynamics, the cardiac drug toxicity testing service founded by UW-Madison stem cell researcher Jamie Thomson. Regina Reynolds, director of corporate communications for EraGen, known for its clinical molecular diagnostics and drug discovery technology, said the BIO conference has been ...

Wisconsin leads in stem cell science, but trails in producing companies

...ormed Cellular Dynamics International to study the toxicity of drugs, particularly those that may be used to treat cardiovascular disease. That company received a combination of loans and grants from the state of Wisconsin, and well as other financing. Why arent more stem cell companies taking root in Wiscon...

Third Wave receives FDA clearance of pharmacogenetic test

...available therapies for colorectal cancer has made toxicity avoidance an important aspect of the clinical decision." The FDA, in its own news release announcing the approval of the Third Wave test, said its use could provide colorectal cancer patients with significant medical benefit. Approximately 150,000 n...

Why are new drugs failing?

...ignificantly curtailed due to its potential cardio toxicity and new label warnings. What about poor Merck? M...tech companies is to identify any potential cardio toxicity before a drug starts clinical trials because most of drug development money is spent at this stage...

Madison-based Mithridion seeks to capitalize on Alzheimer's breakthrough

... it generates another protein that counteracts the toxicity of the first. "So it turns out that this 'good cut' protein from the big parent protein was actually causing this significant induction of these protective mechanism that blocked the bad protein toxicity. That was very interesting," Johnson remarke...

Discovery may halt progression of Alzheimer's

...increase in transthyretin, which was balancing the toxicity of the beta-amyloid, or bad-cut protein." Further experimentation bore out their theory. When Stein introduced an antibody into the mouse brain that prevented transthyretin from binding with beta-amyloid protein, the mice developed early signs of ne...

The birth of biotech: San Francisco, Boston, Geneva or Chicago?

...l time there, Goldwasser worked in the universitys toxicity lab on antidotes for chemical warfare agents (this was after the Japanese attack on Pearl Harbor). Upon graduating, he was drafted and sent to Fort Detrick where he worked on anthrax. He returned to the University of Chicago to complete a doctorate ...

Americas Third Coast nurtures biotechnology

...otency inside targeted cancer cells. The selective toxicity of ribonucleases also makes them ideal for linkage to antibodies or growth factors for therapeutic delivery. We are exploring these kinds of partnerships with antibody companies, says Kauten. Cancer drugs based on ribonucleases look economically fe...

Researchers Push for Faster Drug Development

...bsorption, distribution, metabolism, excretion and toxicity (adme-tox in laboratory parlance). I think that we will focus more on cell-based assays for the future, said Millis, who received her Ph.D. in biochemistry from UW-Madison. It will first of all hopefully decrease the time from drug identification to...