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Method for determining intracapillary solution temperatures: Application to sample zone heating for enhanced fluorescent labeling of proteins
Author(s) -
Giordano Braden C.,
Horsman Katie M.,
Burgi Dean S.,
Ferrance Jerome P.,
Landers James P.
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500633
Subject(s) - reagent , chemistry , fluorescence , chromatography , sample (material) , analytical chemistry (journal) , capillary action , transient (computer programming) , capillary electrophoresis , electrophoresis , sample preparation , materials science , optics , computer science , physics , composite material , operating system
A fundamental premise in CE relies heavily on the assumption that temperature within the capillary is accurately known and controlled. Theoretical calculations for sample zone and BGE temperature during voltage application are presented. We propose that transient elevation of the sample zone temperature allowed for denaturing and renaturing of proteins in the presence of a fluorescent dynamic labeling reagent. Comparison with the extent of labeling possible with standard on‐column dynamic labeling in the absence of elevated temperatures showed order‐of‐magnitude increases in the fluorescence detection sensitivity of proteins with low surface hydrophobicity. As a result, this represents an example where excess heating in the sample zone during electrophoresis can be exploited advantageously.