Navigation Links
Rapid Method Development With the BioLogic DuoFlow Chromatography System for the Purification of His-Tagged Proteins

from crude E. coli lysate on a Profinity IMAC Ni2+-charged resin. In the present study, the concentration of a competitive eluting agent, imidazole, was optimized using the BioLogic DuoFlow system by executing multiple repetitive chromatography runs with a nine-buffer system. The plumbing setup of the system included an SV5-4 select valve (5-port, 4-position) and an AVR9-8 select valve (9-port, 8-position), which were both designed for flexible automated operation. This multiple inlet port configuration, coupled with the buffer scouting feature of the BioLogic DuoFlow software, allowed us to complete this experiment with a single linked scouting run and virtually no user intervention.


Methods
Setup
The column, UV detector, conductivity monitor, BioFrac fraction collector, SV5-4 select valve, AVR7-3 automated sample injection valve, and AVR9-8 select valve were connected to the workstation and controller as illustrated in Figure 1. All nine buffers, with the following compositions, were filtered and connected to the indicated SV5-4 and AVR9-8 valve ports:

A1: loading or equilibration buffer (50 mM potassium phosphate, 50 mM EDTA, 300 mM NaCl, pH 7.5)
A2: cleaning buffer (50 mM potassium acetate, 300 mM NaCl, pH 4.0)
A3: charging buffer (100 mM NiSO4, 300 mM NaCl, pH 4.3)
A4: loading buffer (50 mM potassium phosphate, 300 mM NaCl, pH 8.0)
B1: elution buffer 1 (loading buffer + 75 mM imidazole)
B2: elution buffer 2 (loading buffer + 100 mM imidazole)
B3: elution buffer 3 (loading buffer + 250 mM imidazole)
B6: elution buffer 4 (loading buffer + 50
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
2. A Simple and Rapid Technique to Purify High-Quality Midiprep DNA
3. CastAway Precast Gels for Rapid Automated DNA Sequence Analysis
4. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
5. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
6. Simple and Rapid Isolation of Plant RNA Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit
7. Rapid PCR* Using Active Tube Temperature Control On Thermos PCR Machines
8. Molecular Biology:Easy Dephosphorylation and Rapid DNA Ligation
9. Rapid detection of genomic duplications and deletions using the Agilent 2100 bioanalyzer
10. Rapid hit confirmation and early hERG liability testing using the IonWorks HT system
11. Rapidly Prepare Probes and Markers for Small RNA Analysis
Post Your Comments:
(Date:7/29/2014)... 2014 SoundConnect , an ... is proud to announce the strategic additions of ... their Boston, Charleston and Atlanta offices. SoundConnect’s ... demand for hosted solutions, collaboration and mobility that ... cost. , SoundConnect was founded in 2004, with ...
(Date:7/29/2014)... July 29, 2014 The advanced optical ... brain that are driving a revolution in the neurosciences ... peer-reviewed journal published by SPIE, the international society ... coincides with the launch of the BRAIN Initiative in ... that lay out a vision for and provide examples ...
(Date:7/29/2014)... Cheshire Cat: Can a particle be separated from its ... Communications , published the results of the first Cheshire ... conducted by Chapman University in Orange, CA, and Vienna ... a cat without a grin," thought Alice in Wonderland, ... curious thing I ever saw in my life!" Alice,s ...
(Date:7/29/2014)... July 29, 2014  The Society for Clinical ... updated program for the 9 th annual ... in Amelia Island , Florida.  The focus ... remains unchanged, however, the topics and format will ... opportunity among all stakeholders with a continued focus ...
Breaking Biology Technology:SoundConnect Organization and Culture is Evolving 2SPIE Launches 'Neurophotonics' Journal with Special Section on BRAIN Initiative 2SPIE Launches 'Neurophotonics' Journal with Special Section on BRAIN Initiative 3SPIE Launches 'Neurophotonics' Journal with Special Section on BRAIN Initiative 4The quantum Cheshire cat: Scientists separate a particle from its properties 2The quantum Cheshire cat: Scientists separate a particle from its properties 3The quantum Cheshire cat: Scientists separate a particle from its properties 4TransCelerate, Lilly, Pfizer, and FDA Highlight Collaborative Program at the Site Solutions Summit 2TransCelerate, Lilly, Pfizer, and FDA Highlight Collaborative Program at the Site Solutions Summit 3
... , AUSTIN, Texas, July 27 Patton ... of injection port therapy, announced today that they have entered into ... medical devices used in the treatment of diabetes. , , ... The two companies are working together to design and develop the ...
... SAN DIEGO, July 24 Orexigen Therapeutics, Inc. (Nasdaq: ... exercised their overallotment option to purchase an additional 1,500,000 shares ... public offering that priced on July 23, 2009. Including ... a public offering price of $7.50 per share, resulting in ...
... , , , ... , NES-ZIONA, Israel, July 24 PROLOR Biotech, Inc. (OTC Bulletin ... purchase agreements with private investors to sell up to $10 million of the ... per share. As part of the agreements, the company closed on the ...
Cached Biology Technology:Patton Medical Devices Partners With Unomedical to Develop and Market the i-port Advance(TM) Injection Port for People Taking Multiple Daily Injections 2Patton Medical Devices Partners With Unomedical to Develop and Market the i-port Advance(TM) Injection Port for People Taking Multiple Daily Injections 3Orexigen(R) Therapeutics Announces Full Exercise of Overallotment Option by Underwriters 2PROLOR Biotech Announces $10 Million Preferred Stock Line From Members of The Frost Group 2PROLOR Biotech Announces $10 Million Preferred Stock Line From Members of The Frost Group 3PROLOR Biotech Announces $10 Million Preferred Stock Line From Members of The Frost Group 4
(Date:7/29/2014)... PARIS, 29 JULY 2014 With thirty days to ... for the FEBS-EMBO 2014 Conference are well underway. The ... Biochemical Societies (FEBS), EMBO, and the French Society for ... Saturday, 30 August to Thursday, 4 September at the ... 2014 Conference is the largest meeting for the life ...
(Date:7/29/2014)... study from UC San Francisco is the first to ... overtime and accelerate cellular aging, these negative effects may ... sleeping well. , "The study participants who exercised, ... than the ones who didn,t maintain healthy lifestyles, even ... author Eli Puterman, PhD, assistant professor in the department ...
(Date:7/28/2014)... people in the United States have a circulatory problem ... be painful and may even require surgery in serious ... wasting and, in turn, limb amputation. , At The ... Medical School, scientists tested a non-surgical preventative treatment in ... associated with increased blood circulation. Their proof-of-concept study ...
Breaking Biology News(10 mins):Counting down to FEBS-EMBO 2014 in Paris, France 2Healthy lifestyle may buffer against stress-related cell aging, study says 2Scientists discover genetic switch that can prevent peripheral vascular disease in mice 2
... University of New Hampshire researchers have received a ... farm as a sustainable closed agroecosystem, exploring viable ... grant, from the U.S. Department of Agriculture,s Sustainable ... explore whether closing energy and nutrient cycles could ...
... surface waters in the arid deserts of the U.S. ... fascinated scientists for over two centuries. A new publication ... diversity of closed basins, ancient lake beds, and partially ... Marith C. Reheis of the USGS, "tantalizes scientists with ...
... magazine dedicated to covering the global environment, is launching ... the former editor of Mother Jones and Audubon magazines, ... leading Web sites for commentary and reporting on the ... inaugural edition features articles by noted environmental advocate and ...
Cached Biology News:UNH receives $380,000 grant to study organic dairy as closed ecosystem 2UNH receives $380,000 grant to study organic dairy as closed ecosystem 3Yale Environment 360 magazine launches online 2
BD IMagnet 1 each...
Vertical Rack (five box vapor phase rack), includes 2.0" (5.1cm) boxes...
... Johnstone and M.W. Turner (1997). • ... methods used in the production of ... in immunoassays, the standardisation of antibodies ... including amplification systems, enzyme immunoassays for ...
... Casein Kinase 1, active MW: 34.2kDa ... NaCl, 0.02% Brij-35, 1mM dithiothreitiol with ... data, refer to the Certificate of ... Purification: ammonium salt precipitation followed by ...
Biology Products: