Method for determining intracapillary solution temperatures: application to sample zone heating for enhanced fluorescent labeling of proteins

Electrophoresis
Braden C GiordanoJames P Landers

Abstract

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.

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Citations

Feb 7, 2009·Electrophoresis·Christopher J Evenhuis, Paul R Haddad
Dec 7, 2007·Electrophoresis·Vladislav Dolník
May 13, 2011·Electrophoresis·Claudia Cianciulli, Hermann Wätzig
Feb 7, 2012·Analytica Chimica Acta·Braden C GiordanoAlex Terray
Jul 6, 2014·Pharmacognosy Magazine·Omana Sukumaran Bindhu, Maheshwari Kumari Singh

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