Navigation Links
A Rapid Method for the Purification of Analytical Grade Proteins From Plants Using Preparative SDS-PAGE and Preparative IEF

n using Bio-Rads Econo System low pressure chromatography workstation. Fractions were collected for 5 hours to give a total Model 491 Prep Cell electrophoresis run of 10 hours.

Sample Analysis
Fractions collected from the Model 491 Prep Cell were analyzed by SDS-PAGE using the Mini-Protean II system. Starting with fraction 4, 60 l of every other fraction was mixed with 40 l of sample buffer (125 mM Tris-HCl (6.8), 30% glycerol, 4% b-mercaptoethanol, 4% SDS, and 0.001% bromophenol blue). After being electrophoresed on 7.5% polyacrylamide gels, separated proteins were electrophoretically blotted to PVDF membranes (5.5 x 8.5 cm) at a constant current of 2.0 mA/ cm2 of gel surface for 30 minutes. Blots were either stained with 0.05% Coomassie Blue or probed with antibody developed to tobacco BiP fusion protein (TOBBLP4, gift from Dr. J. Denecke).3

Preparative Isoelectric Focusing
Fractions 35, 36, and 37 from the Model 491 Prep Cell run were pooled and precipitated with 80% AS. The resulting 80% AS pellet was resuspended in 10 ml of dd H2O and dialyzed overnight against the same. The dialyzed sample was then mixed with 40 ml of Rotofor buffer (1% Bio-Lyte 5/7 ampholyte, 1% urea, and 5% glycerol). The sample was focused on the Rotofor cell at a constant power of 10 W for 4 hours at 4 C. Initial voltage was 1,250 and reached the 2,000 volts limit after 1 hour run time. Twenty fractions were collected from the Rotofor cell and each was lyophilized to a final volume of 500 l. Lyophilized samples were analyzed by SDS-PAGE as previously described.


Results
The data in Figu
'"/>

Source:


Page: All 1 2 3 4 5

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:7/3/2008)... 3 /- Cellectis SA today,an...on Pharmaceuticals,Inc. over the use of Cellectis...mbination for the generation of innovative human t... monoclonal antibodies. As part of the,agreement, ...rom Regeneron.,In addition, Regeneron will invest ...
(Date:7/3/2008)...wire-FirstCall/ -- Advanced Life Sciences,Holdings...ny engaged in the,discovery, development and comme...as of infection, cancer and respiratory diseases, ...icrocap(R) Index after the,Russell Investment Grou...global equity indexes on June 27, 2008., (Logo: ...
(Date:7/3/2008)...SEY CITY, New Jersey, July 3,/PRNewswire-FirstCall...ing,developer of microRNA-based diagnostics and th...s Stewart Fourth Annual Growth Conference on,Wedne...tal Hotel in New,York., About Rosetta Genomics, ...he field of microRNA.,Founded in 2000, the company...
(Date:7/3/2008).../- AMDL, Inc. (Amex:,ADL), ...artered in Tustin,Ca., with operations in Shenzhe...as released its investor presentation which is ava...tp://www.amdl.com ., About AMDL:, AMDL, Inc. is ...pany is,headquartered in Tustin, California with o...
Breaking Biology Technology:Cellectis and Regeneron Sign an Agreement Over the use of Homologous Recombination Technology 2Advanced Life Sciences Added to Russell Microcap Index 2Advanced Life Sciences Added to Russell Microcap Index 3AMDL Announces Investor Relations Presentation Available on Website 2
...has passed a bill that allows researchers to work ... will be used against them in a patent dispute. Af...ferent institutions could count as prior art, said... Research Foundation . , ,This could invalidate a ...so far besides making research institutions approa...
...of demostrating Wisconsin,s breakthroughs in biote... Conference closed last Wednesday with a discussi..., ,A panel of out-of-state investors praised the s...ed without capital they wont be able to go further...ent of the Wisconsin Technology Council , said th...
...quire AccuImage Diagnostic Corp. to expand its h... announced on Wednesday. , ,Merge eFilm, the busin...ts to close the deal with the San Francisco-based ...ware, used by radiologists and other specialty phy...tools that can be used for diagnostics of calcium,...
...d it has bought all outstanding stock of Texas-bas...bank transactions and check image processing. , ,M... said in October that it would buy VectorSGIs stoc...to a wholly owned subsidiary. The move followed Me...ns, another company that works on electronic check...
Other Biology Technology:Out-of-state VCs offer advice on biotech investments 2Out-of-state VCs offer advice on biotech investments 3
(Date:7/3/2008)...LE, Fla. --- A farm irrigation canal would seem a ...arket parking lot. , But University of Florida s...udy with wide implications for a longstanding deba...at to amphibians, UF zoologists have found that to...om reproductive system abnormalities than toads ne...
(Date:7/3/2008)...ervation satellites have provided vital informatio...ularly long crisis response window following the d...2 and 3 May 2008. , Immediately after the disast...rian Affairs (OCHA) asked the International Charte...the Charter, for support by providing immediate c...
(Date:7/3/2008)...roblem with antibiotics is that, eventually, bacte...geting the gene that confers such resistance, a ne...ller University scientists tested the new drug, ca...trains of multidrug-resistant Staphylococcus aureu...reat majority of staphylococcal infections worldwi...
(Date:7/2/2008)...s from Hungary, Germany and the U.K. have discover...unds similar to the active ingredient in marijuana...aining healthy skin. This finding on "endocannabi... the October 2008 print issue of, The FASEB Journ...ions ranging from acne to dry skin, and even skin-...
Breaking Biology News(10 mins):New study points to agriculture in frog sexual abnormalities 2New study points to agriculture in frog sexual abnormalities 3Extended cyclone relief efforts aided from space 2Extended cyclone relief efforts aided from space 3New antibiotic beats superbugs at their own game 2New Bill Would Improve Services to Lung Patients 4714 1New Bill Would Improve Services to Lung Patients 4714 2New Bill Would Improve Services to Lung Patients 4714 3Texas Health and Human Services Commission Adopts Mediwares ClosedLoop 28TM 29 Medication Management Strategy 4712 1Texas Health and Human Services Commission Adopts Mediwares ClosedLoop 28TM 29 Medication Management Strategy 4712 2Texas Health and Human Services Commission Adopts Mediwares ClosedLoop 28TM 29 Medication Management Strategy 4712 3Mayo Clinic Calls for Thorough Review of Canadian Pacifics Acquisition of the Dakota Minnesota 26 Eastern Railroad 4710 1Mayo Clinic Calls for Thorough Review of Canadian Pacifics Acquisition of the Dakota Minnesota 26 Eastern Railroad 4710 2Highmark Medicare Services Awarded New Contract from Centers for Medicare 26 Medicaid Services 4708 1Highmark Medicare Services Awarded New Contract from Centers for Medicare 26 Medicaid Services 4708 2
...resulting from dilated cardiomyopathy is rapidly e...ecular mechanisms are being studied and advances h...milial forms. Intracellular [Ca2+]i homeostasis is...ty, and dysregulation of [Ca2+]i is a hallmark of ...ardiac [Ca2+]i transients is calcium entry through...
... for lingering viral infections such as HIV and he...sts it may be possible to deactivate such viruses ..., Princeton scientists Leor Weinberger and Thomas ...s by which human immunodeficiency virus (HIV) and ...eir hosts. The researchers have discovered a speci...
...discovered by Queen’s University researchers may b...esistant bacteria, such as those that cause food p...and activity of the protein ?called YihE or RdoA ?...ents of Biochemistry and Microbiology & Immuno...lopment. , “Our group is the first to solve th...
...ildren,s Hospital in San Francisco have launched a...rocedure using magnets can correct sunken chest, t...same way that orthodontic braces gradually realign...s pectus excavatum, is a deformity of the cartilag...formed cartilage pulls the breastbone inward, maki...
Other Biology News:Protein overexpression at heart of heart failure 2Scientists find potential 'off-switch' for HIV virus 2Scientists find potential 'off-switch' for HIV virus 3Scientists find potential 'off-switch' for HIV virus 4Protein discovery targets antibiotic-resistant bacteria 2Study to explore using magnets to correct 'sunken chest' 2Study to explore using magnets to correct 'sunken chest' 3
a1-Antitrypsin (human, alpha-1 AT)
...osomal vesicles are used to study co-translational...eins. Processing events such as signal peptide cle...glycosylation can be examined by the translation o... these microsomal membranes. In addition, processi...
...tify products ATP-Free using Mo Bio luciferase ass... ATP-Free certification service that will allow yo...vel of accuracy and enable you to deliver them to ...tection limit is 3.5 x 10-12 /l, which will allow ...
...ew Concept in Microarrays: Biomedical Photometrics...p technologies GmbH (Jena, Germany) have collabora...LONDIAG ArrayTube System. The DNAscope AT is base...DNAscope AT automatically reads up to six ArrayTub...
Biology Products: