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

er Stain Plus kit.


Results
Chlorella sorokiniana NADP-specific GDH was purified an additional 8.5-fold for a final 375-fold purification using the Model 491 prep cell (see Table). Nondenaturing preparative electrophoresis of the partially purified NADP-GDH sample was performed under conditions determined to be optimal for analytical native slab gel electrophoresis. A total of 11.5 mg of extremely pure GDH protein was recovered with <2% loss of activity during the procedure. The resolution of the nondenaturing gel in the Model 491 prep cell was sufficient to separate multiple peaks of NADP-GDH activity corresponding to the multiple isoenzymes of the GDH isoenzymes. Analytical SDS-PAGE of the final purified product revealed a single 53 kD band corresponding to the NADP-GDH -subunit and detected no contaminating proteins. The highly purified NADP-GDH protein was recovered in sufficient quantity in its native form to allow NH2- and COOH-terminal sequencing, antigen affinity column production, and the production of hightiter polyclonal and monoclonal antibodies.

These results suggest that the Model 491 prep cell system is amenable to large-scale purification of partially purified proteins in their native, active form. The high resolution of this technique, evidenced by its ability to separate closely migrating isoenzymes, should allow separation of multiple isoenzymes for the purpose of kinetic, biochemical, and immunochemical analyses.


Acknowledgment
We thank Dr LO Ingram for the use of the Model 491 prep cell.


References
Baker AL and Schmidt RR, Intracellular dist
'"/>

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:1/23/2015)... January 23, 2015 Portal Solutions , ... workplaces on Microsoft Office 365 and SharePoint platforms , ... Award program for being one of the region’s fastest-growing mid-sized ... employee growth. , “To be recognized a second year ...
(Date:1/22/2015)... today announces the firm brokered a long-term lease totaling 202,492 ... company, at Two Ledgemont Center in Lexington, Massachusetts ... and Brian Cohen , senior vice president, represented Shire ... Hayden Ave. Photo - http://photos.prnewswire.com/prnh/20150122/170765 ...
(Date:1/22/2015)... San Diego, California (PRWEB) January 22, 2015 ... R Promotional Offer to their comprehensive portfolio of ... Offer allows customers to purchase a Centrifuge 5424/5424 R ... Pipette Bundle of either Eppendorf Research plus or Eppendorf ...
(Date:1/22/2015)... 2015 Diagenode, Inc., a leading ... Bioruptor® and complete solutions for epigenetics research, recently ... alleviating the need for manual processing. The new ... ChIP of histones or transcription factors and a ...
Breaking Biology Technology:Washington SmartCEO Announces Portal Solutions as 2015 Future 50 Award Winner Second Year in a Row 2Washington SmartCEO Announces Portal Solutions as 2015 Future 50 Award Winner Second Year in a Row 3Transwestern?RBJ Advises Shire in 202,000 SF Lease, Creating Boston's Largest Suburban Biotech Campus 2Eppendorf Announces Promotional Bundle Which Includes: Eppendorf Centrifuge 5424, 3-Pack of Pipettes, and Tips - Available Now at Pipette.com 2Diagenode Launches Unique Semi-Automated Solution for Chromatin Immunoprecipitation 2
... Bowman Carney &,Williams, PLLC announces an investigation into ... CGPI ). On February 26, 2008,CollaGenex announced that ... an affiliate of Galderma Pharma S.A. Under the ... per share in cash for each,share owned. This ...
... WILL FOCUS ON COLLECTION, PROCESSING AND STORAGE OF EQUINE ... BONE MARROW AND PLACENTAL TISSUE STEM CELLS, RANCHO ... ), a leading supplier of innovative products and,services that ... its,wholly-owned subsidiary, Vantus, Inc., has signed a formal agreement ...
... 27 NuVasive, Inc. (Nasdaq:,NUVA), a medical device company ... the spine, announced today,that Kevin O,Boyle, Executive Vice President ... Canaccord Adams, Small-Cap Orthopedics,Conference at The Mission Bay Conference ... 4, 2008 at 8:10 a.m. PT., A live ...
Cached Biology Technology:ThermoGenesis Subsidiary Vantus Announces Stem Cell Agreement With UC Davis Center For Equine Health 2ThermoGenesis Subsidiary Vantus Announces Stem Cell Agreement With UC Davis Center For Equine Health 3ThermoGenesis Subsidiary Vantus Announces Stem Cell Agreement With UC Davis Center For Equine Health 4ThermoGenesis Subsidiary Vantus Announces Stem Cell Agreement With UC Davis Center For Equine Health 5NuVasive to Present at Canaccord Adams' Small-Cap Orthopedics Conference 2NuVasive to Present at Canaccord Adams' Small-Cap Orthopedics Conference 3
(Date:12/11/2014)... blood pressure plays a role in human health has been ... for high blood pressure – was first described as a ... used in measuring blood pressure was invented in 1896. ... its triggers and its effects. In fact, recent findings have ...
(Date:12/10/2014)... YORK , Dec. 8, 2014 You,ve been here before: ... can,t remember your password, site key or the answer to your ... teacher? Today, Hoyos Labs , a digital ... put an end to the frustration that comes with usernames, passwords ...
(Date:12/10/2014)... 2014  Wake Forest Baptist Medical Center today announced plans for ... Funding for this $50 million capital project is part of ... summer. The medical education building will be ... complex, adjacent to 525@vine in Wake Forest Innovation Quarter. Construction ...
Breaking Biology News(10 mins):Research points to need for new approaches to treatment of high blood pressure 2Research points to need for new approaches to treatment of high blood pressure 3Research points to need for new approaches to treatment of high blood pressure 4The Password is Finally Dead: Launch of 1U Mobile App Eliminates Need for All Usernames and Passwords 2The Password is Finally Dead: Launch of 1U Mobile App Eliminates Need for All Usernames and Passwords 3Wake Forest Baptist to Build New Medical Education Facility In Wake Forest Innovation Quarter 2Wake Forest Baptist to Build New Medical Education Facility In Wake Forest Innovation Quarter 3Wake Forest Baptist to Build New Medical Education Facility In Wake Forest Innovation Quarter 4
... of researchers from the Department of Biochemistry and Molecular ... Autònoma de Barcelona has discovered that tetanus toxin, which ... against psychological disorders such as depression, anxiety and anorexia, ... as Parkinson's disease. , Tetanus toxin is a neurotoxin ...
... grain sorghum , Trials have begun in Kansas on ... of many plastics, drugs, solvents and food additives. Developed ... form of the bacteria E. coli that metabolizes glucose ... to make key chemical intermediates like succinate is a ...
... scientist from the University of Manchester has discovered the first ... up to 20 million years old. , Two droplets of ... amber which also contains the spider - Filistatidae - a ... The droplets are the first identified examples of spider blood ...
Cached Biology News:Tetanus toxin found to have therapeutic properties 2Bacteria are key to 'green' plastics, drugs 2Bacteria are key to 'green' plastics, drugs 3Spider blood found in 20 million year old fossil 2
Anti-Rat KAPPA/LAMBDA:HRP, Clone MARK-1/MARL-15, Monoclonal Antibody...
MOUSE ANTI RAT KAPPA/LAMBDA:HRP Immunogen: MARK-1: Rat kappa light chains (IR202 and IR968)MARL-15: Rat lambda light chains (RH58, IR31)...
Request Info...
Request Info...
Biology Products: