| Continuous mode of operation for large volume dosing in analytical carrier ampholyte-free isoelectric focusing of proteins applied to off-line detection of fractions. | |
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MedLine Citation:
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PMID: 16922278 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Jana Budilová; Jirí Pazourek; Pavel Krásenský; Jan Pospíchal |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Journal of separation science Volume: 29 ISSN: 1615-9306 ISO Abbreviation: J Sep Sci Publication Date: 2006 Jul |
Date Detail:
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Created Date: 2006-08-22 Completed Date: 2007-08-30 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101088554 Medline TA: J Sep Sci Country: Germany |
Other Details:
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Languages: eng Pagination: 1613-21 Citation Subset: IM |
Affiliation:
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Masaryk University, Department of Analytical Chemistry, Brno, Czech Republic. buda@mail.muni.cz |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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:
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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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