Document Detail


Continuous mode of operation for large volume dosing in analytical carrier ampholyte-free isoelectric focusing of proteins applied to off-line detection of fractions.
MedLine Citation:
PMID:  16922278     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Mass spectrometry is being increasingly used for analysis of proteome complex samples. Sample preparation is often necessary to remove matrix interferences and to concentrate analytes prior to MS measurement. A useful method for this purpose is Carrier Ampholyte Free-Isoelectric Focusing (CAF-IEF). In this paper CAF-IEF of ampholytes was performed on a commercial apparatus EA101 (Villa Labeco, Slovakia) equipped with a specially made column for samples of large volume (up to 0.5 mL). A new continuous mode without voltage interruption or electrolyte replacement was developed. In this mode, a low molecular mass pI marker (PIM 7.4) and low concentrations of myoglobin and insulin (16 mg/L), respectively, were concentrated, and then 5-microL fractions collected for off-line analyses. The total time of focusing was 66 minutes. The concentration of PIM 7.4 in the fractions was increased up to 75 times (determined by UV-VIS spectrometry). The concentration in the fractions was increased up to 30 times for myoglobin and 10 times for insulin.
Authors:
Jana Budilová; Jirí Pazourek; Pavel Krásenský; Jan Pospíchal
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of separation science     Volume:  29     ISSN:  1615-9306     ISO Abbreviation:  J Sep Sci     Publication Date:  2006 Jul 
Date Detail:
Created Date:  2006-08-22     Completed Date:  2007-08-30     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101088554     Medline TA:  J Sep Sci     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  1613-21     Citation Subset:  IM    
Affiliation:
Masaryk University, Department of Analytical Chemistry, Brno, Czech Republic. buda@mail.muni.cz
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MeSH Terms
Descriptor/Qualifier:
Animals
Buffers
Electrophoresis, Capillary / methods
Hydrogen-Ion Concentration
Insulin / analysis
Isoelectric Focusing / instrumentation,  methods*
Myoglobin / analysis
Proteins / analysis*
Proteome / analysis
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods
Spectrophotometry, Ultraviolet
Chemical
Reg. No./Substance:
0/Buffers; 0/Myoglobin; 0/Proteins; 0/Proteome; 11061-68-0/Insulin

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


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