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:9/19/2014)... September 19, 2014 The new , ... , available today on PharmaBoardroom.com for ... crisis that rocked the Spanish economy in 2009, and the ... community has remained pragmatic in times of trouble. ... the fifth biggest European market, and the general conditions to ...
(Date:9/18/2014)... 2014 HealthTronics, Inc ., ... radiology products and services, announced that a poster ... cryoablation) versus external beam radiotherapy (EBRT) is being ... Urological Association (MAAUA) Annual Meeting in Baltimore, Maryland ... of Cryosurgery and External Beam Radiation as Treatments ...
(Date:9/18/2014)... 2014 /PRNewswire-iReach/ -- DeepResearchReports.com adds "2014 Deep Research ... and "2014 Deep Research Report on Global and ... database. Photo - http://photos.prnewswire.com/prnh/20140918/147058 ... Global and China Acetic Acid Industry" is a ... China and Global Acetic Acid Market. ...
(Date:9/18/2014)... Sept. 18, 2014  Health Enhancement Products, Inc. (OTCQB: HEPI), ... algae bioactive compounds and metabolic processes, is pleased to announce ... of directors. Ms. Nola E. Masterson , a ... the board effective September 17, 2014. "We,re ... join our board," states Andrew Dahl , President and ...
Breaking Biology Technology:PharmaBoardroom Releases New Spain Pharmaceuticals Report 2PharmaBoardroom Releases New Spain Pharmaceuticals Report 3HealthTronics Announces Data Comparing Cryotherapy Versus Radiation for Treating Prostate Cancer Poster Presented at the American Urological Association Regional Meeting 2Acetic Acid Market & Sodium Stannate Industry (China, Global) Research Report 2Acetic Acid Market & Sodium Stannate Industry (China, Global) Research Report 3Acetic Acid Market & Sodium Stannate Industry (China, Global) Research Report 4Health Enhancement Announces New Board Member In Advance Of Annual Shareholder Meeting 2
... 10, 2009, NeoStem, Inc. (NYSE Alternext US: NBS) (the ... "Alternext") indicating that the Company is not in compliance ... "Guide"), which requires a listed company to hold meetings ... Company had announced that it planned to hold a ...
... ITMN ) today announced the pricing of an underwritten ... The last reported sale price of InterMune,s common stock on ... $17.96. The offering is expected to close on or ... All of the shares are being sold by the company.UBS ...
... Pharmacyclics, Inc.,(Nasdaq: PCYC ) today reported financial ... net loss for the second quarter of fiscal 2009,was $4.8 ... of $5.9 million,or $0.23 per share, in the second quarter ... expenses were $4.9 million in the second quarter of fiscal,2009, ...
Cached Biology Technology:NeoStem Receives Notice of Noncompliance from NYSE Alternext US 2InterMune Announces Pricing of Public Offering of 3,500,000 Shares of Common Stock 2Pharmacyclics Reports Second Quarter Fiscal 2009 Financial Results 2Pharmacyclics Reports Second Quarter Fiscal 2009 Financial Results 3Pharmacyclics Reports Second Quarter Fiscal 2009 Financial Results 4Pharmacyclics Reports Second Quarter Fiscal 2009 Financial Results 5Pharmacyclics Reports Second Quarter Fiscal 2009 Financial Results 6Pharmacyclics Reports Second Quarter Fiscal 2009 Financial Results 7
(Date:9/21/2014)... warming, are set to rise again in 2014 - reaching ... per cent projected rise in burning fossil fuels is revealed ... UK by researchers at the Tyndall Centre for Climate Change ... of Engineering, Mathematics and Physical Sciences at the University of ... Summit, where world leaders will seek to catalyse action on ...
(Date:9/21/2014)... California, San Diego, have developed a program that predicts ... of genes based on sequences of DNA. They describe ... human embryonic cells in a paper published in ... our cells have the same blueprint, the same DNA, ... author of the report. "Skin cells protect, nerve cells ...
(Date:9/21/2014)... strains of bacteria have emerged that resist even the ... forms of tuberculosis and staphylococcus, infect more than 2 ... the urgent need for new treatments, scientists have discovered ... decade. , MIT engineers have now turned a powerful ... that can disable any target gene, they have shown ...
Breaking Biology News(10 mins):CO2 emissions set to reach new 40 billion ton record high in 2014 2CO2 emissions set to reach new 40 billion ton record high in 2014 3Program predicts placement of chemical tags that control gene activity 2Battling superbugs 2Battling superbugs 3Battling superbugs 4
... from ESA,s Envisat satellite radar are being used to ... Globe solo round-the-world yacht race. CLS, a subsidiary ... new radar applications division (formerly the BOOST Technologies Company), ... Envisat radar imagery to operationally observe oceans at high ...
... seascape that cleaves earlier each springthey may find relief in ... changes in the timing of sea-ice breakup and of snow ... least some polar bears with an alternative source of food. ... "Over 40 years, six subadult male bears were seen among ...
... battle against invading bacteria reaches its peak activity at ... Experiments with the laboratory model organism, Drosophila melanogaster, reveal ... with the body,s circadian rhythm, according to Stanford researchers ... Cell Biology (ASCB) 48th Annual Meeting, Dec. 13-17, 2008 ...
Cached Biology News:Vendee Globe route seen from above 2Goose eggs may help polar bears weather climate change 2Immunity stronger at night than during day 2