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

0 mM imidazole)
B7: elution buffer 5 (loading buffer + 750 mM imidazole)

Experimental
A 1 ml Econo-Column chromatography column (0.5 cm ID x 5 cm) was packed manually with Profinity IMAC Ni2+-charged resin (60 m). Five separate runs were conducted, with either 75, 100, 250, 500, or 750 mM imidazole in the elution buffer. Following elution and a subsequent wash step, the column was regenerated using the loading and charging buffers listed under Setup. These five successive runs were linked through the buffer scouting function of the BioLogic DuoFlow software. For each run, the column was preequilibrated with 10 column volumes (CV) of loading buffer at a flow rate of 1.5 ml/min. Crude E. coli lysate (250 l) containing a 75 kD His-tagged protein was loaded into a 3 ml injection loop and injected into the column through an AVR7-3 valve. Host protein contaminants, which were not retained by the Profinity IMAC resin (Figure 2), were removed by washing the column with 10 CV of the loading buffer. The 75 kD His-tagged protein was then eluted with 5 CV of imidazole elution buffer, followed by an additional wash with 10 CV of the loading buffer to eliminate residual proteins from the column. The column was then regenerated as detailed in the following steps using 10 CV of each buffer: metal removal with the discharging buffer; washing with the cleaning buffer; recharging of the metal ion with the charging buffer; removal of excess metal ions with the cleaning buffer; and finally, reequilibration with the loading buffer.

In order to confirm the separation of the target protein from other E. coli proteins in the crude sample, the peak fractions we
'"/>

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:8/1/2014)... transport of light is the photonic analog of topological ... case, the current flows around the edge of the ... "topological" in that even if electrons encounter impurities in ... losing energy., In the ... regular material but in a meta-material consisting of an ...
(Date:8/1/2014)... , Aug. 1, 2014 Research ... "The World Market for Flow Cytometry in ... http://photos.prnewswire.com/prnh/20130307/600769 ... are now an important component of advanced IVD ... instruments for cancer diagnostics, but their newer clinical ...
(Date:8/1/2014)... One of the major revolutions taking place ... is the practice of using genomic and other molecular ... biology. Personalized diagnostics—laboratory analyses that inform the practice of ... future. This fall, AACC ,s first ever ... this exciting field and the novel technologies driving it ...
(Date:8/1/2014)... 2014 Once a decision has been ... webinar will discuss how to define your tolerance for ... Bob Allen, Senior Director of Facilities Integration at Fluor ... Fluor Industrial Services, and special guest Carrier Li, Director ... Amgen, as they provide an examination of the Conceptual ...
Breaking Biology Technology:On-chip topological light 2On-chip topological light 3The World Market for Flow Cytometry in IVD Applications 2First-Ever AACC Virtual Conference to Focus on Personalized Diagnostics 2Defining Low Cost in Your Solution for Future Facilities, New Webinar for Pharma, Biotech, Medical Device and Healthcare Companies, Hosted by Xtalks 2
... Solos Endoscopy, Inc. (Pink Sheets: SNDY) is pleased ... agreement with Lifeline Biotechnologies, Inc. (Pink Sheets: LLBO) ... to Lifeline Biotechnologies for a combination of stock, ... Endoscopy originally purchased the products from Lifeline Biotechnologies ...
... plc (LSE: SHP, NASDAQ: SHPGY ) announces ... Court of the Southern District of,New York against Mylan ... for infringement of Shire,s,patent: U.S. Patent No. 5,968,976 (",976 ... filed in response to an Abbreviated New Drug Application,("ANDA") ...
... 20 A new, innovative study,shows that a ... plus dexamethasone not only prevents chemotherapy-induced nausea,and vomiting ... oncology,patients. Data presented today at the ESMO Symposium ... New data presented today at ...
Cached Biology Technology:Solos Endoscopy, Inc. Sells Ovascope and Mastascope Product Line to Lifeline Biotechnologies, Inc. 2Shire Files Lawsuit Against Mylan for Infringement of FOSRENOL(R) (Lanthanum Carbonate) Patent 2Shire Files Lawsuit Against Mylan for Infringement of FOSRENOL(R) (Lanthanum Carbonate) Patent 3Palonosetron Warrants Adequate Caloric Intake in Oncology Patients Receiving High Emetogenic Chemotherapy 2Palonosetron Warrants Adequate Caloric Intake in Oncology Patients Receiving High Emetogenic Chemotherapy 3Palonosetron Warrants Adequate Caloric Intake in Oncology Patients Receiving High Emetogenic Chemotherapy 4
(Date:7/31/2014)... available in German . ... gases similar to a sponge that soaks up liquids. ... hydrogen or greenhouse gases. However, loading of many MOFs ... Technology (KIT) now report in Nature Communications ... MOF surface. This can be prevented by water-free synthesis ...
(Date:7/31/2014)... the photovoltaic effect in 1839, humankind has sought to ... its own purposes. In a new research report published ... FASEB Journal , scientists may have uncovered a ... focusing on a naturally occurring combination of lipids that ... over time and across speciessuggests that this specific natural ...
(Date:7/31/2014)... Professor of Clinical Surgery at LSU Health Sciences Center ... team of plastic and reconstructive surgeons who report a ... select patients for a specific surgical migraine treatment technique. ... surgery to decompress the nerves that trigger migraines experienced ... The study, which confirms the benefit of surgical treatment ...
Breaking Biology News(10 mins):Free pores for molecule transport 2Scientists shine bright new light on how living things capture energy from the sun 2Surgeons report significant migraine relief from cosmetic eyelid surgery technique 2
... disease ever eradicated. In two studies in the Dec. 15 ... scientists are working to ensure that once it is gone, ... immune systems were weakened by HIV would re-introduce poliovirus into ... switching forms of vaccine from a live, attenuated vaccine to ...
... for millennia, but there has been great controversy about ... Proceedings of the National Academies of Sciences, a team ... University of Wisconsin-Madison has for the first time demonstrated ... potato. , The scientists analyzed DNA markers in 261 ...
... Pumps implanted into the chest to maintain circulation can ... end-stage heart failure who are not candidates for heart ... led by Duke University Medical Center cardiologists. , The ... employed when the heart's left ventricle ?the chamber of ...
Cached Biology News:Polio vaccination strategies assessed as eradication nears 2Polio vaccination strategies assessed as eradication nears 3Finding rewrites the evolutionary history of the origin of potatoes 2Implantable pumps extend lives of patients too sick for transplant 2Implantable pumps extend lives of patients too sick for transplant 3
... Thermo has established an expertise in ... both for solid support synthesis and synthesis ... controlled by Mass Spectrometry and analytical HPLC ... provide products that meet the highest quality ...
The Wellwash 4 Mk 2 microplate washer combines flexibility, simplicity and ease of use. This reliable microplate washer meets the exacting standards of today's ELISA assay requirements....
... These handy dispensers save space and ... and other garb. They are the ... parts used in labs, processing and ... charges and the particles they attract. ...
... dispensers save space and organize ... and other garb. They are ... for small parts used in ... acrylic. Many other sizes and ...
Biology Products: