Navigation Links
Preparative Nondenaturing Gel Electrophoresis to Purify NADP-Specific Glutamate Dehydrogenase From Chlorella, Rev B

ced in 14,000 MW cutoff dialysis tubing. The concentrated sample was dialyzed at 4C against 28.8 mM Tris, 192 mM glycine, 2 mM DTT (pH 8.4) for 30 min. The dialyzed enzyme preparation was clarified by centrifugation at 20,000 x g for 10 min and was combined with 3 ml of 40% sucrose and 1 ml of 0.02% Bromophenol Blue.

Preparative Nondenaturing Gel Electrophoresis
For preparative nondenaturing gel electrophoresis, a 3 cm high 7% acrylamide (28 acrylamide:0.735 bis-acrylamide, pH 8.8) resolving gel and a 2 cm high 2% acrylamide (1.6 acrylamide:0.4 bis-acrylamide, pH 6.6) stacking gel were cast in the 28 mm ID gel tube of the Model 491 prep cell. The resolving gel was polymerized in 374 mM Tris (pH 8.8) using 140 g/ml ammonium persulfate and 1.12 l/ml TEMED. The stacking gel was polymerized in 39 mM Tris (pH 6.6), 12.7% sucrose using 0.07% riboflavin-5-phosphate (Bio-Rad) and 0.365 l/ml TEMED. Stacking gel polymerization was achieved by exposure to two 5 W fluorescent lights for 45 min. Both gels were cooled as per the Model 491 prep cell instruction manual during the polymerization process. All acrylamide stock solutions were pretreated with AG 501-X8 mixed bed resin to remove any contaminating acrylic acid to prevent in vitro acylation of proteins during the electrophoresis process. The preparative gel was pre-electrophoresed for 10 min at 15 mA constant power in upper/lower gel electrophoresis buffer containing 28.8 mM Tris, 192 mM glycine (pH 8.4, 4C). The elution buffer reservoir was filled with elution buffer comprised of 28.8 mM Tris, 192 mM glycine, 2 mM DTT, and 0.1 mM NADP+ (pH 8.4, 4C). The entire Model 491 prep cell was cooled to 4C during the electrophoresis process
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
2. Detection of Platinum Species in Plant Material by Preparative Isotachophoresis Using the Model 491 Prep Cell, Rev A
3. Preparative Nondenaturing Gel Electrophoresis Used in the Purification of an Esterase Involved in Insecticide Resistance, Rev C
4. Preparative SDS-PAGE Electrophoresis of a Recombinant Epstein-Barr Virus Encoded Protein and Its Application in Serodiagnostic Test Systems, Rev C
5. Purification and Characterization of beta-Lactoglobulin Genetic Variants A and B Using Preparative Electrophoresis and Isoelectric Focusing
6. A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF
7. Semi Preparative Purification of Proteins
8. A Versatile Power Supply for All Electrophoresis Applications
9. A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis
10. Continuous-Elution Electrophoresis for Purification of the Baculovirus-Expressed Coronavirus Structural Proteins, Rev A
11. Detection of K-ras Point Mutations in the Pancreas by Constant Denaturing Gel Electrophoresis Using the DCode System
Post Your Comments:
(Date:7/24/2014)... July 24, 2014 Gain recognition ... Supply Chain of the biotech industry. Nominations are ... Awards, brought to you by the Bio ... of supply chain management professionals for the past ... stakeholders of the Biotech industry – Manufacturers, Service ...
(Date:7/24/2014)... MENLO PARK, Calif. , July 24, 2014 ... biotechnology company in the emerging field of regenerative ... live webcast of a presentation to investors on ... 1:30 p.m. PDT.  The presentation will include an ... pipeline. To access the webcast, visit http://www.ustream.tv/channel/asterias-biotherapeutics ...
(Date:7/24/2014)... equipment, trained personnel, and detection dogs to safeguard ... A revolutionary new electronic chip with nano-sized chemical ... easier. , The groundbreaking nanotechnology-inspired sensor, devised by ... School of Chemistry and Center for Nanoscience and ... picks up the scent of explosives molecules better ...
(Date:7/24/2014)... Yet, scientists are making progress in devising suitable ... approach relies on quantum dotsa kind of artificial ... A new study demonstrates that changing the coupling ... electrical impulses can help better control them. This ... as quantum information units, which would produce faster ...
Breaking Biology Technology:Call for Submission: BSMA’s Supply Chain Management Innovation Awards 2Call for Submission: BSMA’s Supply Chain Management Innovation Awards 3Asterias Biotherapeutics Announces Live Investor Webcast 2Asterias Biotherapeutics Announces Live Investor Webcast 3Asterias Biotherapeutics Announces Live Investor Webcast 4Nano-sized chip 'sniffs out' explosives far better than trained dogs 2
... a largely unseen battle now raging in the workplace. It ... are managers who covet the traditional employment deal money, ... commitment to playing the organizational game by traditional rules, e.g., ... unquestioning willingness to do whatever the business requires. Only Type ...
... University of Wisconsin-Madison leads the nation in the number ... in 2006. UW-Madison's total this year is up to ... ,The foundation bestows the awards on young researchers in ... biology, neuroscience and physics. Sloan Fellowships carry a two-year, ...
... Defense Agency, an information security consulting firm headquartered here ... been chosen to challenge eight student teams in an ... complexity and sophistication, so it's essential to support collegiate ... of cyber defenders," said Sami Saydjari, president and founder ...
Cached Biology Technology:Who will win the battle to redefine the workplace? 2Who will win the battle to redefine the workplace? 3Who will win the battle to redefine the workplace? 4Who will win the battle to redefine the workplace? 5UW-Madison has 7 Sloan research fellows, top in nation 2
(Date:7/25/2014)... MA A study published online in the ... a novel approach to preventing cervical cancer based on ... cancer after removal of a discrete population of cells ... a study that looked at squamocolumnar junction cells, or ... and have been implicated as the origins of cervical ...
(Date:7/25/2014)... professor of biomedical engineering at the University of ... engineering and artificial organ development., "Introduction to Tissue ... to entering into the field of artificial organ ... biomedical engineering at UH, served as a series ... there are other published books on the subject ...
(Date:7/25/2014)... has opened the way for the development of new lines ... annual barley production is second only to wheat with 7-8 ... most important diseases of barley. , Senior Research Scientist Dr ... growths on the cell walls of barley plants that block ... research, by the ARC Centre of Excellence in Plant Cell ...
Breaking Biology News(10 mins):Clearing cells to prevent cervical cancer 2University of Houston researcher publishes textbook on tissue engineering 2New hope for powdery mildew resistant barley 2
... from the atmosphere are unlikely to offer an economically feasible ... to a report issued by the American Physical Society and ... not kid ourselves that we can pour all the carbon ... it out later at little cost," said Socolow, a professor ...
... scientists have discovered how two closely related proteins guide ... and repelling these axons as they navigate the most ... make remarkably precise connections. The discovery, reported April ... proteins belonging to the "semaphorin" family of guidance cues ...
... of Botany (KIB), part of the Chinese Academy of ... Natural Products and Bioprospecting (NPAB). As a ... in the SpringerOpen portfolio, available on www.springerlink.com . ... Science (CLoS), a collection comprising more than 90 English-language ...
Cached Biology News:Report: Direct removal of carbon dioxide from air likely not viable 2Johns Hopkins scientists reveal nerve cells' navigation system 2Johns Hopkins scientists reveal nerve cells' navigation system 3New SpringerOpen journal from the Chinese Academy of Sciences 2