Method for determining intracapillary solution temperatures: application to sample zone heating for enhanced fluorescent labeling of proteins.
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.
Giordano
Braden C
BC
Department of Chemistry, University of Virginia, Charlottesville, VA 22904, USA.
Horsman
Katie M
KM
Burgi
Dean S
DS
Ferrance
Jerome P
JP
Landers
James P
JP
D019075:Electrophoresis, Capillary / Q000592:standards*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006361:Heating; D011489:Protein Denaturation; D011506:Proteins / Q000737:chemistry*; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013696:Temperature
D005456:Fluorescent Dyes; D011506:Proteins; D012996:Solutions