Navigation Links
Whole Gel Eluter Purification of a Functional Multiprotein DNA Replication Complex, Rev A

on, polyethylene glycol precipitation, ion-exchange chromatography, and density gradient sedimentation. The integrity of the DNA synthesome has been shown to be maintained after treatment with detergents, salt, RNase, DNase, chromatography on DEAEcellulose (Whatman) and Q-Sepharose (Pharmacia), and following sedimentation in sucrose and glycerol density gradients, indicating that the ready co-purification of the proteins with one another was independent of nonspecific interaction with other cellular macromolecular components.24 Native polyacrylamide gel electrophoresis (PAGE) of the synthesome from HeLa cells revealed the presence of several high molecular weight multiprotein species.7 One of these complexes was readily recognized in western blot analysis by a monoclonal antibody against the DNA replication essential protein DNA polymerase α.7 This DNA polymerase α containing complex was shown to have a high specific in vitro simian virus 40 (SV40) origin dependent DNA replication activity. We have recently found that Bio-Rads Whole Gel Eluter can greatly aid in the purification of the DNA synthesome. What follows is the description of the conditions we developed for the isolation of the DNA synthesome using the Whole Gel Eluter.


Materials and Methods
CELL CULTURE
Suspension cultures of HeLa cells were grown in Jokliks modified Eagles medium supplemented with 5% each of calf and fetal bovine serum. Exponentially growing cells were harvested and washed three times with phosphate-buffered saline (PBS). The cells were then pelleted by low-speed centrifugation. The cell pellets were stored at -80 C prior to initiating subcellular
'"/>

Source:


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

Related biology technology :

1. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
2. Protocol for GenomePlex Whole Genome Amplification from Buccal Swab
3. Protocol for GenomePlex Whole Genome Amplification from Blood Card
4. Protocol for GenomePlex Whole Genome Amplification from Plant
5. Protocol for GenomePlex Whole Genome Amplification from Soil
6. Protocol for GenomePlex Whole Genome Amplification from Whole Blood
7. Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue
8. Bacterial Whole Genome Array Analysis
9. RNAi: Get the Whole Story
10. Total RNA from Whole Blood for Expression Profiling
11. Fractionation of Bovicola ovis Homogenates Using the Mini Whole Gel Eluter
Post Your Comments:
(Date:8/19/2014)... Pa. , Aug. 19, 2014 A ... has been nominated for a Centers for Disease Control ... addressing a major public health concern. ... System, is one of a team of researchers nominated ... Charles C. Shepard Science Award for Data Methods and ...
(Date:8/19/2014)... XenoTech announces an exclusive partnership ... production from bioactive small molecules. Based on ... enables convenient and timely assessment of critical metabolite ... President of Commercial Operations, Christian Darabant, “the value ... can be measured directly by comparison with the ...
(Date:8/19/2014)... 2014   Synthetic Biologics, Inc. (NYSE ... biologic and drug programs targeting specific pathogens that ... its novel C. difficile development program ... Interscience Conference on Antimicrobial Agents and Chemotherapy (ICAAC), ... Synthetic Biologics, Senior Vice President, ...
(Date:8/18/2014)... 2014) -- A new therapy developed by researchers ... College of Medicine and Columbia University Medical Center ... interventional cardiovascular disease treatment. , The researchers ... molecular therapy could selectively inhibit blood vessel re-narrowing ... procedure using a balloon catheter to open narrowed ...
Breaking Biology Technology:Geisinger investigator nominated for federal award 2Geisinger investigator nominated for federal award 3Beyond Metabolite Identification, Testable Quantities Now Offered by XenoTech 2Beyond Metabolite Identification, Testable Quantities Now Offered by XenoTech 3Synthetic Biologics Announces Late-Breaking Poster Presentation for C. difficile Program at 54th ICAAC 2Synthetic Biologics Announces Late-Breaking Poster Presentation for C. difficile Program at 54th ICAAC 3Synthetic Biologics Announces Late-Breaking Poster Presentation for C. difficile Program at 54th ICAAC 4Selective therapy may improve artery repair after interventional cardiovascular procedures 2Selective therapy may improve artery repair after interventional cardiovascular procedures 3
... HEIGHTS, N.J., Feb. 16, 2011 Zymes LLC is ... two-year $476,000 grant from the Michael J. Fox Foundation,s ... titled, "Evaluation of the Neuroprotectivity Ability of Zymes, ... disease): Preclinical Validation and Dose Optimizations for Clinical Study" ...
... Pa., Feb. 16, 2011 Tengion, Inc. (Nasdaq: ... it has been engaged in negotiations with a publicly traded ... merger.  As a result of the recent increases in the ... acquirer has terminated merger negotiations. As has ...
... An important milestone toward the realization of a large-scale quantum ... quantum control of light, were accomplished by a team of ... The study, published in the Feb. 7 issue of ... Zhejiang University, China, and NEC Corporation, Japan. The experimental effort ...
Cached Biology Technology:Michael J. Fox Foundation Awards Grant to Zymes LLC 2Tengion Addresses Recent Trading Activity in its Common Stock 2International team of scientists says it's high 'NOON' for microwave photons 2
(Date:8/20/2014)... MA Using population-based screening outcomes of approximately 3 ... including four researchers at the University of Massachusetts Medical ... immunodeficiency (SCID) can be successfully implemented across public health ... published in the Aug. 20 issue of the ... ) showed the rate of SCID in newborns is ...
(Date:8/20/2014)... suffered concussions ready to return to action? A new ... athletes who head back on the field with medical ... their abilities to simultaneously walk and do simple mental ... their balance and/or altered walking speed, was found in ... returned to activity in less than a month. Seven ...
(Date:8/20/2014)... Your chairs, synthetic rugs and plastic bags could one ... waste rather than petroleum, scientists are now reporting. The ... help the world deal with its agricultural and plastic ... . , Athanassia Athanassiou, Ilker S. Bayer and colleagues ... plastic,s popularity is constantly growing. In 2012, its production ...
Breaking Biology News(10 mins):Newborn screening expansion offers early diagnosis and treatment to infants with SCID 2Newborn screening expansion offers early diagnosis and treatment to infants with SCID 3Recovery reversal seen in Oregon study of returning concussed athletes 2Recovery reversal seen in Oregon study of returning concussed athletes 3
... the Faculty of Veterinary Medicine at Universitat Autnoma ... Research in Agricultural Genomics (CRAG), the Centre for ... for Agrarian Technology and Research in Madrid and ... the first partial genome sequencing of an Iberian ...
... The debate surrounding carbon capture and storage intensifies as ... Berkeley National Laboratory (Berkeley Lab) examine the capacity for ... in the new journal Greenhouse Gases: Science & ... the conclusions drawn in an earlier study by Ehlig-Economides ...
... biochemistry is how proteins recognize "correct" interaction partners in ... critical to know than in the brain, where interactions ... A team of biologists at the Salk Institute for ... regulates availability of a brain channel that modulates neuronal ...
Cached Biology News:First partial sequencing of an Iberian pig 2Carbon capture and storage: Carbon dioxide pressure dissipates in underground reservoirs 2Salk scientists crack molecular code regulating neuronal excitability 2Salk scientists crack molecular code regulating neuronal excitability 3
... coli strain containing an overproducing clone of ... Transcriptase catalyzes the polymerization of DNA using ... copies of large mRNAs, >10 kb, may ... much lower RNase H activity than AMV ...
... coli containing a clone of the human Topoisomerase ... II alters the topological state of nucleic acids ... transient break which generates a separate DNA helix ... DNA passage mechanism, the enzyme can relax negatively ...
... Type Liquid. In 100 mM KCl, ... EDTA, 20% glycerol, pH 7.9. AVOID FREEZE/THAW CYCLES. ... monomeric protein of 765 amino acids encoded by ... both positive and negative supercoiled DNAs without the ...
Request Info...
Biology Products: