Navigation Links
Quantitative Analysis of Protein-DNA Associations in Vivo Using Real-Time PCR, Rev A

Swarup K Chakrabarti and Raghavendra G Mirmira*, Departments of Medicine and Pharmacology, Division of Endocrinology and Metabolism, University of Virginia Health Sciences Center, 450 Ray C Hunt Dr, Box 801407, Charlottesville, VA 22903 USA

*Corresponding author


Introduction
The chromatin immunoprecipitation (ChIP) assay has been used in recent years to identify the association of histone complexes and transcription factors with specific genomic DNA targets in cells of Drosophila or yeast. As shown in Figure 1, the technique first involves the treatment of cells with formaldehyde or ultraviolet light to covalently crosslink proteins to DNA. The chromatin from the cells is then isolated and fragmented (by restriction digestion or sonication), and the protein of interest is immunoprecipitated using a specific antibody. The coimmunoprecipitated DNA fragments are then liberated from the protein by treatment with proteinase K, and the DNA fragments are identified by Southern blotting or PCR (Kuo and Allis 1999). More recently, studies have established the utility of the ChIP assay in studying histone complex associations and transcription factor binding in mammalian cells as well (Boyd and Farnham 1999). However, a potential barrier to the quantitative study of some mammalian transcription factors by the ChIP assay is their relatively low level of expression in cells, coupled with the rather large size of the mammalian genome compared to yeast or Drosophila. Southern blot detection of DNA does not offer the necessary sensitivity for such studies, nor is traditional PCR easily amenable to quantitation. However, we demonstrate here that real-time PCR wit
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. Mx4000 Multiplex Quantitative PCR System*,PPP
2. Improved Quantitative Selectivity of Clenbuterol in Human Urine using High Resolution on the TSQ Quantum Mass Spectrometer
3. The LightCycler is the only System for Quantitative PCR that is as Flexible as You are!
4. Quantitative Measurement of Cell Proliferation Using the BrdU ELISA: A Comparison Between Colorimetric and Chemiluminescent Detection
5. Quantitative end-point RT-PCR gene expression measurement using the Agilent 2100 Bioanalyzer and standardized RT-PCR
6. Detection of Genetically Modified Soybean in Processed Foods Using Real-Time Quantitative PCR with SYBR Green I Dye on the DNA Engine Opticon 2 System
7. Qualitative and Quantitative Assessment of Plasmid DNA Isolation, Rev A
8. Rapid, Reproducible Real-Time Quantitative RT-PCR Using the iCycler iQ Real-Time PCR Detection System and iQ Supermix, Rev A
9. Quantitative Analysis of GFP in Plant Extracts Using the VersaFluor Fluorometer System
10. Multiplexed Quantitative Peptide Assays for Protein Biomarkers of Cardiovascular Disease in Human Plasma
11. Quantitative Analysis of the Cell Cycle in Imaging Applications
Post Your Comments:
(Date:12/19/2014)... -- Relmada Therapeutics, Inc. (OTCQB: RLMD), a clinical-stage company ... announced the results of its annual general meeting of ... year in Relmada,s history and it set the stage ... Sergio Traversa , chief executive officer of Relmada. "Over ... human resource and clinical development objectives that position Relmada ...
(Date:12/17/2014)... 2014 Achim Noack has been named ... Group. In this position, Noack will assist Jerry Stoller, ... develop marketing strategy and supervise global marketing management and ... has tremendous knowledge and experience in the crop protection ... experience and innovative thinking will be a great asset ...
(Date:12/17/2014)... IN (PRWEB) December 17, 2014 ... Laboratory Excellence following its most recent COLA inspection. ... by COLA, a national healthcare accreditation organization. Accreditation ... standards of quality in day-to-day operations, demonstrate continued ... pass a rigorous on-site laboratory survey. ...
(Date:12/17/2014)... 17, 2014  United Therapeutics Corporation (NASDAQ: ... well as changes to Martine Rothblatt , Ph.D,s ... United Therapeutics announced the promotion of ... Officer and David Zaccardelli , Pharm.D. to Executive ... these promotions, Dr. Rothblatt,s title will change from Chairman ...
Breaking Biology Technology:Relmada Therapeutics Announces Results of Annual Shareholders Meeting 2Achim Noack Joins Stoller Group as Global Vice President of Marketing and Administration 2Xeno Diagnostics Receives 3rd Award for Laboratory Excellence 2United Therapeutics Corporation Announces Executive Promotions and CEO Compensation Change 2United Therapeutics Corporation Announces Executive Promotions and CEO Compensation Change 3United Therapeutics Corporation Announces Executive Promotions and CEO Compensation Change 4
... PARIS , April 27, 2010 ... - French ... Inactivation of,Protein-based Infectious Agents Linked to Fatal Brain Diseases , ... Advanced Sterilization Products (ASP) announced today ...
... , ... partners with RURO to offer GLP/GMP certified hosting services. , ... Annapolis, MD (Vocus) April 27, 2010 -- ... with laboratory software provider RURO. RURO’s leading edge laboratory applications can now be ...
... calculations are helping physicists gain molecular level insight ... which change shape under the influence magnetic fields. ... compact alternatives to conventional motors and actuators. But ... exhibit much larger changes in shape than most ...
Cached Biology Technology:ASP Sterrad Technology Approved by AFSSAPS for Total Inactivation of Prions 2ASP Sterrad Technology Approved by AFSSAPS for Total Inactivation of Prions 3ASP Sterrad Technology Approved by AFSSAPS for Total Inactivation of Prions 4Sidus BioData Partners with Life Science Software Provider RuRo 2
(Date:12/19/2014)... , Dec. 18, 2014 Research and Markets ... addition of the "iPhone 5S Fingerprint Sensor ... report to their offering. ... acquisition of AuthenTec in July 2012, Apple introduced ... It is currently the only device of Apple ...
(Date:12/17/2014)... --Research and Markets ( http://www.researchandmarkets.com/research/s6qzt5/samsung_galaxy_s5 ) ... Galaxy S5 - Home Button Synaptics Fingerprint Sensor ... http://photos.prnewswire.com/prnh/20130307/600769 ... the iPhone 5S, Samsung introduces for the first ... The Galaxy S5 home button presents an ...
(Date:12/17/2014)... 16, 2014 Research and Markets ... the "Global Chemical Sensor Market 2015-2019" ... http://photos.prnewswire.com/prnh/20130307/600769 One major trend ... for medical sensors in biomedical applications. Chemical sensors ... and correct diagnosis during surgical procedures. The Global ...
Breaking Biology News(10 mins):iPhone 5S Fingerprint Sensor - Apple/AuthenTec TMDR92 & Sapphire - Technology Report 2Samsung Galaxy S5 - Home Button Synaptics Fingerprint Sensor Technology Analysis 2Global Chemical Sensor Market 2015-2019: Key Vendors are Abbott Laboratories, Bayer, Hoffmann La-Roche, Johnson & Johnson, NGK Spark Plugs and Robert Bosch 2
... today announced the recipients of their 2009 Communication Awards. ... these prestigious awards recognize excellence in reporting and communicating ... With support from the W.M. Keck Foundation, the National ... This year,s winners will be honored during a ceremony ...
... Society of Interventional Radiology presents Catheter Lysis ... featuring top interventional radiologists, neurointerventionists, neurologists and ... and/or upgrade skills to incorporate acute ischemic ... The course incorporates lectures, panel discussions, teaching ...
... reporting discovery of a much sought after crack in the ... one-sixth of the world,s population, causing stomach ulcers and other ... key chemical pathway that the bacteria need for survival. Their ... fight these hard-to-treat microbes, is scheduled for the October 16 ...
Cached Biology News:Academies announce winners of 2009 Communication Awards 2Academies announce winners of 2009 Communication Awards 3Top interventional radiologists, neurointerventionists explore stroke care through CLOTS 2