Document Detail


Use of quasi-isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performances.
MedLine Citation:
PMID:  18383017     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The use of quasi-isoelectric anolytes and catholytes has been investigated to improve CIEF performances. Narrow pH cuts of carrier ampholytes (NC) have been compared to more conventional couples of anolytes/catholytes (phosphoric acid/sodium hydroxide and glutamic acid/lysine). First, a CIEF setup that consists in a bare silica capillary and 70:30 water/glycerol separation medium has been used. The experiments have shown that when using NC instead of more classical anolytes and catholytes, an increase in the protein detection time was observed and the resolutions obtained for neutral and acidic proteins were doubled. Moreover, according to the NC fraction used, the resolution was modified. In order to investigate further the mechanisms involved, a second setup using a capillary coated with hydroxypropylcellulose was used. With this setup no difference has been observed when changing anolyte and catholyte nature. A simple methodology has then been developed to evaluate EOF during focusing and mobilization steps of CIEF experiments. It highlighted the crucial role played by EOF when using a bare silica capillary. EOF indeed decreased by 33% during mobilization step when using NC instead of classical anolytes and catholytes.
Authors:
Martine Poitevin; Gabriel Peltre; Stephanie Descroix
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Electrophoresis     Volume:  29     ISSN:  0173-0835     ISO Abbreviation:  Electrophoresis     Publication Date:  2008 Apr 
Date Detail:
Created Date:  2008-04-23     Completed Date:  2008-08-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8204476     Medline TA:  Electrophoresis     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1687-93     Citation Subset:  IM    
Affiliation:
Laboratoire Environnement et Chimie Analytique, UMR CNRS 7121, ESPCI, Paris, France.
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MeSH Terms
Descriptor/Qualifier:
Buffers
Cellulose / analogs & derivatives,  chemistry
Hydrogen-Ion Concentration
Isoelectric Focusing / methods*
Chemical
Reg. No./Substance:
0/Buffers; 9004-34-6/Cellulose; 9004-64-2/hydroxypropylcellulose

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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