Navigation Links
Separation and Comparison of Proteins From Virulent and Nonvirulent Strains of the Fish Pathogen Flavobacterium psychrophilum, Using a 2-D Electrophoretic Approach

les), and were vortexed between cycles for 12 min. Endonuclease was added to a concentration of 150 U/ml and samples were incubated at room temperature for 30 min to digest DNA. Samples were then centrifuged (12,000 x g) and supernatant containing hydrophilic proteins was retained (E1). To separate hydrophobic proteins, reagent 2 (8 M urea, 4% CHAPS, 40 mM Tris, and 0.2% Bio-Lyte 3/10 carrier ampholytes) containing 2 mM tributylphosphine (TBP) was added to the pellet. The sample was vortexed for 2 min and centrifuged as above. The supernatant, containing hydrophobic proteins (corresponding primarily to loosely bound membrane proteins), was retained (E2). Reagent 3 (5 M urea, 2 M thiourea, 2% CHAPS, 2% SB 3-10, 40 mM Tris and 0.2% Bio-Lyte 3/10 carrier ampholytes) with 2 mM TBP was then added to the pellet and the process repeated to obtain highly hydrophobic proteins generally associated with tightly bound membrane proteins (E3). Samples were either subjected to isoelectric focusing immediately following extraction or stored at -80C until needed.

Biotinylation of Cell Surface Components
In order to detect cell surface membrane proteins and allow comparison to sequentially extracted proteins, viable bacteria were biotinylated prior to protein extraction. Bacteria were harvested in log-phase growth at a concentration of approximately 5.0 x 107 viable bacteria/ml (0.4 absorbance) and washed twice in ice-cold PBS (pH 8.0) taking care not to disrupt or lyse cells. The bacteria were resuspended to an absorbance of 0.2 at 525 nm and 1.7 mg of sulfo-NHS-LC-biotin was added per ml of bacteria. This mixture was allowed to rock for 30 min at room temperature. The bacteria were then wa
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Separation Reproducibility With the BioLogic Chromatography Systems
2. Separation of 16S rRNA PCR Products From a Model Community Using Temporal Temperature Gradient Gel Electrophoresis, Rev A
3. Separation of HLA-DRB Alleles by Denaturing Gradient Gel Electrophoresis using the DCode System
4. Separation and Quantitation of R and S Enantiomers using Normal Phase Chromatography on an API 3000 LC/MS/MS System
5. BD IMag Human Regulatory T Lymphocyte Separation Set for Isolation of Treg Cells
6. Separation of Peptides by Hydrophilic Interaction Chromatography Using TSKgel Amide-80
7. Unique Surface Treatment of Novel TSK-GEL ODS Columns Provide Separation of Aqueous and Fat Soluble Vitamins
8. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases
9. Performance Comparisons of Commercial RT-PCR Systems
10. A Comparison Study of Lipid Tranfection Reagents in A549, NIH 3T3 and COS-7 Cell Lines
11. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
Post Your Comments:
(Date:2/27/2015)... CA (PRWEB) February 27, 2015 Cytokinetics, ... forces on February 28 with patients and health care ... awareness for Rare Disease Day®. Rare Disease Day ... and calling attention to the special challenges faced by ... them. , “Cytokinetics is proud to stand alongside patients ...
(Date:2/27/2015)...   PureTech , a science and technology R&D ... and technologies in the healthcare sector, announced today the ... Member of the Board of Directors of Sanofi, to ... been a pleasure to know Chris for many years ... working together more closely now," said Dr. Robert ...
(Date:2/26/2015)... Feb. 26, 2015 On behalf of biotech ... companies, Wilentz, Goldman & Spitzer, P.A. (Wilentz) ... against Genewiz Inc. in excess of $10 million. The ... trade secrets, and that it improperly hired one or ... denying motions for judgment notwithstanding the verdict, a new ...
(Date:2/26/2015)... - SQI Diagnostics Inc. ("SQI" or the "Company") (TSX-V: ... and commercializes proprietary technologies and products for advanced microarray ... the fiscal first quarter ended December  31, 2014. All ... (CAD), unless otherwise stated. "During the ... our existing customers and add to our sales pipeline," ...
Breaking Biology Technology:Cytokinetics Joins Global Movement to Raise Awareness for Rare Diseases 2Cytokinetics Joins Global Movement to Raise Awareness for Rare Diseases 3Cytokinetics Joins Global Movement to Raise Awareness for Rare Diseases 4Cytokinetics Joins Global Movement to Raise Awareness for Rare Diseases 5Cytokinetics Joins Global Movement to Raise Awareness for Rare Diseases 6Christopher Viehbacher, former CEO of Sanofi, Joins PureTech's Board of Directors 2Christopher Viehbacher, former CEO of Sanofi, Joins PureTech's Board of Directors 3Christopher Viehbacher, former CEO of Sanofi, Joins PureTech's Board of Directors 4Wilentz Client GenScript Awarded $10 Million in Trade Secrets Case Against Genewiz 2Wilentz Client GenScript Awarded $10 Million in Trade Secrets Case Against Genewiz 3SQI Diagnostics Reports First Quarter Financial Results 2SQI Diagnostics Reports First Quarter Financial Results 3SQI Diagnostics Reports First Quarter Financial Results 4SQI Diagnostics Reports First Quarter Financial Results 5SQI Diagnostics Reports First Quarter Financial Results 6
... In response to recent press coverage concerning investment ... Minnesota, Burrill & Company would like to clarify that ... have been in active discussions for several months with ... project," said G. Steven Burrill, CEO, Burrill & Company. ...
... National Ergonomics Conference and Exposition (ErgoExpo) at Caesars Palace, ... on the importance of ergonomics, safety and employee wellness ... topics of program management and return on investment, the ... specifically for those working in offices, industrial, lab, healthcare, ...
... Sept. 16 Regado Biosciences, a privately held company ... agents, announced that David J. Mazzo, PhD, President and ... Sciences Conference.  Dr. Mazzo,s presentation will be given at ... Wednesday, September 22, 2010 at 8:00 a.m. EDT in ...
Cached Biology Technology:Burrill & Company Responds to Recent Press Coverage About Biopark Investment at Elk Run 22010 National Ergonomics Conference Coming To Las Vegas, Nov. 30 - Dec. 3 With Larger Expo; 22010 National Ergonomics Conference Coming To Las Vegas, Nov. 30 - Dec. 3 With Larger Expo; 32010 National Ergonomics Conference Coming To Las Vegas, Nov. 30 - Dec. 3 With Larger Expo; 4Regado Biosciences to Present at the UBS Global Life Sciences Conference on September 22, 2010 in New York City 2Regado Biosciences to Present at the UBS Global Life Sciences Conference on September 22, 2010 in New York City 3
(Date:2/5/2015)... Feb. 5, 2015  Marken is starting its 35 ... has launched a new marketing campaign to solidify its ... The new campaign focuses on First as ... shipments. The first headline in the series, ... priorities with its client,s priorities. Marken recognizes the need ...
(Date:2/5/2015)... NXT-ID, Inc. (NASDAQ: NXTD ... company focused on the growing mobile commerce market, announces ... CEO Conference on January 29 th at 10:30 ... CEO of NXT-ID will present and host a live ... wallet and its full suite of biometric technologies. Investors ...
(Date:1/22/2015)... 2015 Valid S.A. ("Company") (BM&FBOVESPA: VLID3 ON), is pleased ... State of Washington,s Department of Licensing, ... and central issuance system for driver,s licenses and identification cards ... planning and development will start in January 2015, with enrollment ...
Breaking Biology News(10 mins):Marken Launches New Patient-Centric Campaign 2CEO of NXT-ID, Inc. to Discuss Company's Biometric Technologies at RedChip Online Conference January 29th 2CEO of NXT-ID, Inc. to Discuss Company's Biometric Technologies at RedChip Online Conference January 29th 3Valid USA Signs Contract For Washington Driver's License Issuance 2
... The initial results of the first computer model ... representation of individual clouds have been analyzed by ... Research Center (IPRC) at the University of Hawai`i ... Technology (JAMSTEC), and the University of Tokyo. ...
... learn how to do something by trying it ourselves may ... in neuroscience. It turns out that humans, primates, some birds, ... in the same pattern whether performing or just observing a ... learning motor tasks involving hand eye coordination, and possibly also ...
... Tufts University,s School of Engineering has been selected ... the field of catalysis, particularly for new insights ... fuel conversion and pollutant processes. Election ... members by their peers. Flytzani-Stephanopoulos, professor in the ...
Cached Biology News:How mirror neurons allow us to learn and socialize by going through the motions in the head 2Tufts University Prof. Maria Flytzani-Stephanopoulos named as AAAS Fellow 2